Drawell Gentier 96E User Manual
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Drawell Gentier 96E User Manual

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User Manual
Please read the manual before installation and operation.
Drawell International Technology Limited
Chongqing Drawell Instrument CO,.Ltd
Add:Suite 2705,Building No.12,Shiyou Road No.1,Yuzhong District, Chongqing,China
Tel: 0086-023-63268643
Shanghai Drawell Scientific Instrument Co.,Ltd
Add : Suite 1117,Lane561 XiuChuan Rd.,PuDong New Area,Shanghai,China
Web : www.drawell.com.cn
Email : sales05@drawell.com.cn

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  • Page 1 Please read the manual before installation and operation. Drawell International Technology Limited Chongqing Drawell Instrument CO,.Ltd Add:Suite 2705,Building No.12,Shiyou Road No.1,Yuzhong District, Chongqing,China Tel: 0086-023-63268643 Shanghai Drawell Scientific Instrument Co.,Ltd Add : Suite 1117,Lane561 XiuChuan Rd.,PuDong New Area,Shanghai,China Web : www.drawell.com.cn Email : sales05@drawell.com.cn...
  • Page 2 Gentier Real-Time PCR System Gentier Real-Time PCR System User Manual Drawell International Technology Limited...
  • Page 3 Gentier Real-Time PCR System User Manual Version: V1.8 User Manual Revision Time: 08-Apr - 2019 Suitable Instrument Model: Gentier 96E, Gentier 96S, Gentier 96R, Gentier 96C Medical Device Manufacturing Enterprise License: Shaanxi FDA Production License No. 20170023 is a registered trademark for Drawell International Technology Limited...
  • Page 4 Gentier Real-Time PCR System Safty Labels on Transport Package Label Description Fragile: the items inside are fragile, please handle with care. This Side Up: indicates the upward side ofthe transport package. Keep Dry: keep the transport package away from rain or any liquid. IVD Instrument: the product belongs to In Vitro Diagnostic equipment.
  • Page 5 Gentier Real-Time PCR System Symbols Used in This Manual Symbol Description Caution: reminding uesrs to pay attention to a certain operation. Operating the Gentier instrument in any manner unspecified in this manual may results in instrument damage or abnormal function. Reminding: providing important informations regarding the Gentier instrument operation and successful application, including the information explained in further detail elsewhere in this manual.
  • Page 6 Gentier Real-Time PCR System Safety and Regulatory Compliance The operation, maintenance and repair of Gentier instrument shall strictly follow the safety specifications listed in this section and through this manual. The design of Gentier instrument has fully considered its biological contamination protection, electrical safety protection and mechanical motion protection.
  • Page 7 Gentier Real-Time PCR System Caution: Gentier instrument installation and transportation should be perfomed by professional engineer or under professional guidelines. Caution: please remove the transport lock after instrument installation and reserve the transport lock for future use. Caution: please confirm you have removed the transport lock before power on the instrument.
  • Page 8 Gentier Real-Time PCR System Reminding: under normal circumstances, please use the instrument attached power cord. If the original power cord is destroyed, please substitute it with an equal one. Caution: for protection against electric shock hazards, the Gentier instrument must be grounded properly.
  • Page 9 Gentier Real-Time PCR System Reminding: the working environment of the Gentier instrument should be under normal atmosphere (the altitude should below 2000m). V. Biological Safety and Precautions Biohazard: the sample objects of the Gentier instrument are nucleic acids, please consider all samples are with potential biohazard, and please take applicable safety protection measures and wear appropriate protective goggles, clothing and gloves while processing the samples.

  • Page 10: Table Of Contents

    Gentier Real-Time PCR System Content 1. A ........................... 1 A. OVERVIEW............................1 PPLICATION IELDS 2. P ................1 RODUCT ONSTITUTE AND NSTRUMENT TRUCTURE 2.1 Product Constitute........................1 3. P ........................2 2.2 Instrument Structure......................... 1 RODUCT NTRODUCTION 3.1.1 Instrument General Parameters..................2 3.1 Instrument Parameters and Characters...................2 3.1.2 Instrument Technical Parameters..................4 3.1.3 Instrument Characters......................
  • Page 11 Gentier Real-Time PCR System 1. S ....................18 D. APPLICATION SOFTWARE OPERATION..................18 TARTUP PPLICATION OFTWARE 2. Q ..........................19 UICK TART 3. M ..........................22 NTERFACE 3.1 Menu Bar..........................23 3.2 Tool Bar............................35 3.3.1 Run Setting........................38 3.3 Operation Area.........................36 3.3.2 Sample Setting.........................
  • Page 12 Gentier Real-Time PCR System 1. I ................. 120 F. INSTRUMENT CLEANING AND MAINTENANCE............... 120 NSTRUMENT LEANING PERATION NSTRUCTIONS 1.1 Clean Instrument Shell......................120 1.2 Clean Touch Screen........................121 2. I ..............122 1.3 Clean Loading Platform and Sample Block................121 NSTRUMENT AINTENANCE PERATION NSTRUCTIONS...

  • Page 13: Overview

    Gentier Real-Time PCR System A. Overview 1. Application Fields The Gentier real-time PCR system is intended for performing rapid, accurate polymerase chain reaction (PCR), meanwhile real-time measuring the nucleic acid signals from DNA-binding fluorescent dyes or labeled probes and converts them to comparative quantitative readouts of DNA or reverse transcribed RNA.

  • Page 14: Roduct Introduction

    Gentier Real-Time PCR System Figure A-1. Gentier Instrument Stucture 2. Sample block 1. Touch screen 3. Loading platform 6. Air vent 4&5. Cooling fan 7. Power switch 9. Power socket 8. Fuse tube box 10&12. USB interface 11. Internet access 13.
  • Page 15 Gentier Real-Time PCR System ▶ Package Specification: Dimension: 690mm(L) × 580mm (W) × 720mm (H); Weight: 42kg; ▶ Voltage: AC 100~240V; Power Specification: Frequency: 50~60Hz; Rated power: 900VA; ▶ less than 3 meters; Power Cord: ▶ Cat 5e FTP cable with length less than 3 meters; Network Cable: ▶...

  • Page 16: Instrument Technical Parameters

    Gentier Real-Time PCR System 3.1.2 Instrument Technical Parameters Thermal Parameters ▶ Temperature Accuracy: ≤ 0.1°C; ▶ Temperature Uniformity: ± 0.1°C; ▶ Temperature Precision: ≤ 0.1°C; ▶ Max Heating and Cooling Ramp: 6.1°C/s Optical Parameters ▶ Excitation Light Source: LED light sources; ▶...
  • Page 17 Gentier Real-Time PCR System Man-machine Interactive System Parameters Touch Screen: with built in 10.4’ inches color touch screen, providing more simple ▶ operation. The instrument can run alone without the control computer. The stand-alone running saves more space and energy. USB interface: the USB interface of Gentier instrument is only used to connect USB disk, ▶...

  • Page 18: Instrument Characters

    Gentier Real-Time PCR System 3.1.3 Instrument Characters ▶ Touch Screen Operation: Gentier instrument possesses a built-in 10.4’-inches colored touch screen, very easy for operation. It can run independently from the computer; ▶ Independent Temperature Control: Gentier instrument adopts independent temperature control technology and high repeatability temperature zone.

  • Page 19: Reagent Specifications

    Gentier Real-Time PCR System ▶ Multiple Functions: the software possesses multiple analysis functions that can adapt to a variety of experimental requirements. Such as absolute quantification analysis, relative quantification analysis, melting curve analysis, high resolution melting (HRM) analysis, genotyping analysis, end point fluorescence analysis, etc. ▶...

  • Page 20: Gentier Instrument Installation

    Gentier Real-Time PCR System B. Gentier Instrument Installation 1. Unpacking Instructions The Gentier transport package is as shown in figure B-1. The Gentier instrument and its accensories are well preserved in a carton case. In order to prevent the collision and oscillation during transportation, the Gentier instrument is sealed with plastic bag and well supported by protective foams.
  • Page 21 Gentier Real-Time PCR System S tep: please hold the Gentier instrument on both sides, take it out of the carton and place it on steady plane, shown in figure B-2e, B-2f. Figure B-2. Unpacking Steps Reminding: in order to prevent the formation of condensation water, please do not open the transport package until it reaches the room temperature.

  • Page 22: Orking Nvironmental Equirements

    Gentier Real-Time PCR System Reminding: please check the package integrity before open it. In case there is any damage, collision or water mark, please contact the transport department or our company. Reminding: please check the instrument and accessories according to the packing list and ensure that all components are present and intact, report any missing items to XATL Co., LTD.

  • Page 23: Instrument Power Requirements

    Gentier Real-Time PCR System For protection against overheating hazards, the openings on the Gentier instrument are designed for ventilation. Please do not block or cover these ventilation openings while it is running. Do not place the Gentier instrument on soft surface, the instrument base may sink into the soft surface and block up the air inlet beneath.

  • Page 24: Nstrument Installation

    Gentier Real-Time PCR System Reminding: please check the power cord periodically, substitute with an equal one in case it is found to be damaged. 3. Instrument Installation 3.1 Remove Transport Lock After unpacking, please do not directly power on the Gentier instrument, it is fixed by transport lock during transportation.
  • Page 25 Gentier Real-Time PCR System Figure B 3. Remove transport lock...

  • Page 26: External Device Connection

    Gentier Real-Time PCR System 3.2 External Device Connection Please plug the corresponding power cords of Gentier instrument, control computer and printer to the power supply respectively. Then please confirm the connections of external devices, such as the control computer, display, keypad, mouse and printer, etc. The external device connnection is completed.
  • Page 27 Gentier Real-Time PCR System Figure B 4a. Network information window default network information Figure B 4b. Control computer network setting IP address: the first 3 octets of control computer IP adress should be identical to the Gentier instrument IP address. Subnet mask: the control computer subnet mask should be identical to the Gentier instrument subnet mask.

  • Page 28: The Preparation Before Experiment

    Gentier Real-Time PCR System C. The Preparation before Experiment 1. Instrument Self Inspection The Gentier instrument possesses self-inspection function, it is recommended to let the instrument conduct the self-inspection before running any experiment, to ensure that it can work normally. S tep: switch on the power switch of Gentier instrument.
  • Page 29 Gentier Real-Time PCR System place the cosumables that contain the sample and PCR reagent mixture on the sample block and press the < Open/Close > function key again to withdraw S tep: the loading platform. select or edit an experiment program and then start running. S tep...

  • Page 30: Application Software Operation

    Among all the 96 throughputs Gentier instrument models, Gentier 96E instrument model possesses the most comprehensive functions. Therefore, the Gentier 96E instrument...

  • Page 31: Quick Start Bar

    Gentier Real-Time PCR System After starting the application software, the Welcome screen will automatically switch to the startup interface and pop up quick start bar on this interface, as shown in figure D-2. Figure D - 2. Startup interface of application software 2.
  • Page 32 Gentier Real-Time PCR System Figure D 2.1 Quick start bar ntroduction & Parameter Descriptions Quick Start Bar I The current User Name is displayed on the top of quick start bar and user can click < Switch User > to change the current user account. Login: user can click <...
  • Page 33 Gentier Real-Time PCR System Figure D-2.1b Quick start bar - Add User Quick Start tab: includes four shortcut keys: < New Experiment >, < New Experimentfrom ExistedExperiment >, < Open Data File > and < InstrumentManagement>. Click < New Experiment > and the application software will automatically pop up the new experiment dialog box, as shown in figure D 2.1c.

  • Page 34: Main Interface

    Gentier Real-Time PCR System Click < Instrument Management > and the application software will automatically pop up the instrument management interface; user can manage all instruments within the local area network (LAN). For operation details, please refer to D-3.1 Menu Bar - Tool (T) submenu- InstrumentManagement.

  • Page 35: Menu Bar

    Gentier Real-Time PCR System Figure D 3. Main Interface 3.1 Menu Bar The menu bar of application software includes: File (F), View (V), Tool (T), Option (O) and Help (H) five submenus. File (F) submenu: includes nine options. Options and Functional Description New Experiment (N): create a new experiment.
  • Page 36 Gentier Real-Time PCR System Open Data File...: user can select an experiment data file from the save path and click < Open > to view and analysis the experiment data file. Recent Files: user can click the experiment name from the list to open the selected data file. Close Experiment: close an opened or created experiment file.
  • Page 37 Gentier Real-Time PCR System On-line: display the connection status between the application software and the instrument: indicates disconnected status; indicates connected status; Loading Platform: display the status of Loading Platform, which is Close or Open; Status: display the running state of the connected instrument, which is Ready or Running; Tool (T) submenu: include three options.
  • Page 38 Gentier Real-Time PCR System Instrument List: displays relevant information of instrument connected, including Model, Name, Serial No., Remarks, IP Address and whether the instrument is set as the Default Instrument. There are four function keys under the list: < Add >, < Edit >, < Delete > and < Set as Default Instrument >.
  • Page 39 Gentier Real-Time PCR System c. Click < Delete > to delete the instrument selected in the list. d. Select any instrument in the Instrument List and click < Set as Default Instrument > to set it as the default instrument, it will be automatically connected after startup of application software.
  • Page 40 Gentier Real-Time PCR System File Transmission tab: to transmit the file between the control computer and the connected Gentier instrument, as shown in figure D-3.2d. Figure D-3.2d Instrument Management interface - File Transmission tab a. Local Directory: user can select a local experiment file directory or experiment data file directory from the drop down list of Local Directory.
  • Page 41 Gentier Real-Time PCR System Figure D - 3.3 Tool submenu - T Calculator interface Interface Introduction and Functional Description User can click four keys to input the forward primer sequence in Forward Primer input box; User can click four keys to input the reverse primer sequence in Reverse Primer input box;...
  • Page 42 Gentier Real-Time PCR System Figure D-3.4 Research Report interface - Research Report Interface Introduction and Functional Description ▶ Report Information Selection Bar: user can check the relevant information check boxes in the report information selection bar to determine the information to be displayed and printed in research report.
  • Page 43 Gentier Real-Time PCR System Option (O) submenu: includes three options. Options and Functional Description Configuration Management: Click Configuration Management option in the Option submenu and the application software will automatically pop up the configuration management interface, as shown in figure D-3.5a. Figure D-3.5a Option submenu - Configuration Management interface - Default Path tab Interface Introduction and Functional Description ▶...
  • Page 44 Gentier Real-Time PCR System User can check Do you want to use the last saved position check box to determine whether to use the same default file path for saving experiment file or template next time. If the Default File Path is successfully set and user want to save the experiment file to another path, please click File >...
  • Page 45 Gentier Real-Time PCR System User can check the PANA connection check box in the PANA label and fill in the IP address box below to ask for the IP address of the PANA full-automatic nucleic acid workstation to be connected. Finally, click test to check whether the PANA instrument can be connected, , as shown in figure D-3.5c.
  • Page 46 English: check English option and the software language will switch to English after restarting the application sofware. Help (H) submenu: includes three options. Options and Functional Description User Manual: open the Gentier Real- Time PCR System User Manual. Home Page: visit the home page ofXi'an Drawell Science and Technology Co., Ltd.

  • Page 47: Tool Bar

    Gentier Real-Time PCR System About: display the application software version and copyright information, as shown in figure D-3.7. Figure D - 3.7 Help submenu - About interface 3.2 Tool Bar The Tool Bar of the application software consists of eight commonly used function icons. Options and Functional Description New Experiment >: create a new empty experiment.

  • Page 48: Operation Area

    Gentier Real-Time PCR System QuickStart Bar. 3.3 Operation Area After creating a new experiment file or open an experiment file, the operation area on the main interface of the application software will be activated, as shown in figure D-4. Figure D-4. Operation area on main interface The experiment file created or opened will be displayed in the experiment file tab at the bottom of operation area and user can select or close any experiment file in this tab.
  • Page 49 Gentier Real-Time PCR System Reminding: when user select the different tab in the operation area, different function icons will be added accordingly in the tool bar of application software. Run Setting tab: when the Run Setting tab is selected on the main interface, two additional function icons will be shown in the tool bar on the run setting interface.

  • Page 50: Run Setting

    Gentier Real-Time PCR System 3.3.1 Run Setting The operation area on main interface displays Run Setting tab by default, as shown in figure D-5. Figure D 5. Run Setting interface Run Setting The Run Setting interface consists five parts: temperature program area, stage and step lists, experiment and step editing areas.
  • Page 51 Gentier Real-Time PCR System Figure D-5.1 Temperature program area – Example temperature program Introduction to Temperature Program Area Stages are longitudinal separated; if a stage contains several steps, these steps are separated by solid blue line; The corresponding stage type and cycle number are displayed below each stage box and the corresponding step number is displayed above the step box;...
  • Page 52 Gentier Real-Time PCR System Figure D 5.2 Experiment editing area Tube Type: user can choose the consumable type of current experiment from drop-down ▶ list of Tube Type, including Clear, White and Frosted. Reaction Volume: user can enter the reaction volume for current experiment in the ▶...
  • Page 53 Gentier Real-Time PCR System Reminding: user can also double click the cycle number below the stage box in the temperature program area to complete the cycle setting. Reminding: the cycle number setting range is 1-99. User can add or delete any stage according to the experiment requirements or adjust the ▶...
  • Page 54 Gentier Real-Time PCR System Introduction to Stage Type Window ▶ The application software provides the following stage types: Preincubation, Reverse and Cooling. User can choose the corresponding predefined stage in the stage type window Transcription, 2 Step Amplification, 3 Step Amplification, Melting, Continuous Melting and click <Add >...
  • Page 55 Introduction to Step Setting Dialog box Step Mode: user can set the step mode for the selected step in the drop-down list of Step Mode, which including Standard, Touchdown, Long and Gradient. Model Gentier 96E Gentier 96S Gentier 96R Gentier 96C...
  • Page 56 Gentier Real-Time PCR System Temperature: the temperature in °C, which is to be held for a defined time. User can use ▶ keys, or manually enter the current step temperature in the Temperature input box. Reminding: the temperature setting range is 0.0°C -100.0°C. Time: the time for which the temperature is to be held.
  • Page 57 Gentier Real-Time PCR System Reminding: the initial and target temperature setting range is 0.0 °C - 100.0 °C. Reminding: the start cycle setting range is 1-max cycle number ofthe current stage. Reminding: the Delta Temperature setting range is 0.1°C- 5.0°C. III.
  • Page 58 Gentier Real-Time PCR System Figure D-5.6d Step Setting Dialog Box – Gradient Step Mode Reminding: the gradient step mode is not available on the application sofware of Gentier 96S and Gentier 96C instrument models. Reminding: the temperature center setting range is 35.5°C - 99.5°C. Reminding: the temperature offset setting range is 0.5°C - 20.0°C.
  • Page 59 Gentier Real-Time PCR System Melting: melting stage allows the system to read the fluorescence signals after each temperature increment. If the current step is the last step of melting stage, user can set the temperature Increment in the Step Setting dialog box for the current step, as shown in figure 5.6f.

  • Page 60: Sample Setting

    Gentier Real-Time PCR System 3.3.2 Sample Setting After finishing the experiment run setting, user can click Sample Setting tab to enter the sample setting interface, which is consisted of sample setting area and sample property editing area, as shown in figure D-6. Figure D - 6.
  • Page 61 Gentier Real-Time PCR System Figure D-6.1 Sample Setting interface - Sample Setting area Introduction to Sample Setting Area ▶ User can select the relevant sample well in the sample setting area and operations as follows: Click and select a single well in sample setting area to. Press [Ctrl] key on computer keyboard and click corresponding wells to select multiple discontinuous wells;...
  • Page 62 Dye: user can check the relevant dye check box to determine the detection dyes for the selected wells. Reminding: figure D-6.2 takes Gentier 96E/96S type instrument models as an example. The application software displays the corresponding dye of six channels; and the Gentier 96R/96C instrument models will display the corresponding dye of four channels.
  • Page 63 Gentier Real-Time PCR System Reminding: user can click the color block before the Gene input box to select a data analysis color for all samples with the current gene name. Standard Setting: if the sample type for the selected well is Standard, the Standard ▶...
  • Page 64 Gentier Real-Time PCR System Figure D-6.4 Sample Setting interface - Auto Concentration Setting Window User can set the starting concentration for a series of diluted standard samples in the Starting Conc. input box; User can select the concentration dilution factor for a series of diluted standard samples from Dilution Factor drop-down list;...
  • Page 65 Gentier Real-Time PCR System ▶ Replicate: according to experiment requirements, user can classfy the same samples into one replicate group in the Replicate group setting box, as shown in figure D-6.5. Figure D-6.5 Sample Setting interface - Replicate a. Replicate No.: user can select the corresponding sample wells in the sample setting area and select the replicate group number from the Replicate No.
  • Page 66 Gentier Real-Time PCR System Click < OK > and the application software will automatically divide the replicate group for all the selected samples; User can click < Cancel > and the application software will give up the current replicate setting and automatically return to the sample setting interface. Sample ID : ▶...

  • Page 67: Run Monitoring

    Gentier Real-Time PCR System Figure D-6.7 Sample Setting interface - QC Failure Condition Setting User can manually enter or use keys to set the QC failure conditions, and click < OK> to confirm the setting. User can click < Default > to clear all QC failure condition settings. ▶...
  • Page 68 Gentier Real-Time PCR System Figure D 7. Run Monitoring interface Interface Introduction and Parameter Description The run monitoring interface consists of six functional modules: Real-time Fluorescence, Temperature Program, Sample Setting, Run Info, Sample Info and Heat Map; after entering the run monitoring interface. This interface is divided into four areas and the application software displays four functional modules (Real-time Fluorescence, Temperature Program, Sample Setting, Run Info) by default.
  • Page 69 Gentier Real-Time PCR System Figure D-7.1 Run Monitoring interface - Real-time Fluorescence function module ▶ User can set fluorescence curve display mode from the drop-down list on the bottom left of Real-time Fluorescence functional module. The real-time fluorescence curve can be displayed according to Well Color, Dye Color, Sample Color and Gene Color.
  • Page 70 Gentier Real-Time PCR System ▶ Export >: user can click < Export > icon to export the current real-time fluorescence monitoring diagram. < 2. Temperature Program: display the temperature program of the current running experiment, as shown in figure D-7.2 Figure D-7.2 Run Monitoring interface - Temperature Program function module ▶...
  • Page 71 Gentier Real-Time PCR System ▶ Parameter introduction: same with the parameters introduced in D-3.3.2 Sample Setting - Sample Setting area. ▶ Function key introduction: user can select one or more sample wells in the sample setting function module to display the corresponding fluorescence curve and relevant data in the Real-time Fluorescence and Sample Info function modules.

  • Page 72: Analysis

    Gentier Real-Time PCR System Figure D - 7.5 Run Monitoring interface - Sample Info function module Heat Map: display the real time fluorescence heat map of the current running experiment, as shown in figure D 7.6. Figure D-7.6 Run Monitoring interface - Heat Map function module 3.3.4 Analysis After experiment running, user can click the Analysis tab to enter the analysis interface.

  • Page 73: Abs Quant

    Gentier Real-Time PCR System Figure D-8. New Analysis window ▶ The application software provides user with six analysis methods: Abs Quant, Rel Quant, Melting Curve, High Resolution Melting, Genotyping and End PointFluorescence. Select Stage: user can choose the stages that need to be analyzed. Select Step: user can choose the step that need to be analyzed.
  • Page 74 Gentier Real-Time PCR System modules (Amplification Curve, Standard Curve, Sample Setting, Result Table) by default, as shown in figure D-9. Reminding: user can select the functional module displayed on the analysis interface from the drop-down list on the top right corner of each area. Reminding: user can click the <...
  • Page 75 Gentier Real-Time PCR System Figure D-9.1 Abs Quant analysis interface - Amplification Curve functional module ▶ Parameter introduction: X axis represents the cycle number of the current experiment; Y axis represents the Relative Fluorescence Unit (RFU); ▶ Function key introduction: User can select the amplification curve display mode from the drop down list on the bottom left of Amplification Curve functional module.
  • Page 76 Gentier Real-Time PCR System Figure D- 9.1a Abs Quant analysis interface - Coordinate Range Setting ▶ Export >: user can click < Export > icon to export the current amplification curve diagram. < ▶ Log View >: user can click < Log View >...
  • Page 77 Gentier Real-Time PCR System ▶ Parameter introduction: X axis represents the log concentration of samples; Y axis represents the threshold Ct value; Dye: display the relevant dye of standard curve; Target Gene: display the target gene of standard curve; Slope: display the slope of standard curve; Eficiency: display the amplification efficiency of standard curve;...
  • Page 78 Gentier Real-Time PCR System file, then import it to the standard curve list. The corresponding standard curve diagram will be displayed in the Standard Curve Preview area below. b. Select Standard Curve: user can call one or more standard curve from the selected standard curve file according to the current experiment requirements.
  • Page 79 Gentier Real-Time PCR System Figure D-9.3 Abs Quant analysis interface - Sample Setting functional module IV. Result Table: display the sample details and result data of the current experiment. The result table functional module consists of Result and Statistics sub-tabs. The the Result sub-tab is selected by default on the result table functional module, which display the sample details and results of the current experiment, as shown in figure D-9.4a.
  • Page 80 Gentier Real-Time PCR System User can click Statistics sub-tab to view the statistical results of the current experiment, as shown in figure D-9.4b. Figure D - 9.4b Result Table functional module – Statistics sub - tab Raw Curve: display the diagram of raw amplification curve without subtract baseline, as shown in figure D-9.5.
  • Page 81 Gentier Real-Time PCR System Figure D-9.6 Abs Quant analysis interface - Raw Fluorescence functional module ▶ X axis shows channel 1-6 (Gentier 96E/96S) or channel 1-4 (Gentier 96R/96C). Parameter introduction: Y axis shows the relative fluorescence unit ((RFU). Different colors represent A-H rows of sample wells.
  • Page 82 Gentier Real-Time PCR System Ct: user can check Ct option and click any dye to view the relevant Ct heat map. Concentration: user can select Concentration option and click any dye to view the relevant concentration heat map. Fluorescence: user can select Fluorescence option and select the cycle number from drop- down list;...
  • Page 83 Gentier Real-Time PCR System ▶ Analysis Mode: user can check the analysis mode check box of Reference Dye and Baseline Gain Calibration according to the experiment requirements, and set the corresponding parameters in the Baseline list. a. Baseline Gain Calibration: in general conditions, the application software select the Baseline Gain Calibration analysis mode by default, which provides user with Automatic Baseline and Manual Baseline two baseline setting methods.
  • Page 84 Gentier Real-Time PCR System Gene and Sample tab: user can set the dye, gene and sample required for the analysis of the current experiment within the gene and sample tab, as shown in figure D-10.2. Figure D-10.2 Abs Quant Analysis Setting interface – Gene and Sample tab ▶...

  • Page 85: Rel Quant

    Gentier Real-Time PCR System 3.3.4.2 Rel Quant The relative quantification experiment aims to compare the expression level of two or more genes within a same individual. Usually, endogenous housekeeping genes with conctant expression level are taken as the internal reference genes. The relative quantification is realized by measuring the changes of the target genes expression level related to the internal reference genes.
  • Page 86 Gentier Real-Time PCR System are same with those introduced in D-3.3.4.1Abs Quant. Bar Chart: dispaly the relative ratio relationship between target gene & sample and reference gene & sample, as shown in figure D-11.1. Figure D-11.1 Rel Quant analysis interface - Bar Chart function module ▶...
  • Page 87 Gentier Real-Time PCR System Figure D 11.2 Rel Quant interface Display Setting window a. Y axis: If user select Linear option, the Bar Chart functional module will display the linear coordinates of Y axis; if user select Logarithm option, the Bar Chart functional module will display the logarithm coordinates of Y axis.
  • Page 88 Gentier Real-Time PCR System analysis of the current experiment within the gene and sample tab, as shown in figure D - 12. Figure D 12 Rel Quant Analysis Setting interface Gene and Sample tab ▶ Dye and Gene: display the name oftarget gene and its marking dye. a.

  • Page 89: Melting Curve

    Gentier Real-Time PCR System quantification analysis interface. 3.3.4.3 Melting Curve For intercalating dyes (such as SYBR Green) and non cleavable hybridization probes, the fluorescence intensity is proportional to the amount of double-strand DNA (dsDNA). However, these dsDNA including specific and non-specific PCR products, which means that the presence of primer dimers and other non specific products can affect the quality of real time PCR data.
  • Page 90 Gentier Real-Time PCR System Figure D-13. Melting Curve analysis interface Interface Introduction and Parameter Description Reminding: the parameter descriptions of Sample Setting, Result Table and Heat Map functional modules are same with those introduced in D-3.3.4.1 Abs Quant. Melting Curve: display the fluorescence intensity against the temperature, as shown in figure D-13.1.
  • Page 91 Gentier Real-Time PCR System curve diagram, or click the right mouse button to zoom out. Melting Peak: display the first negative derivative of the fluorescence intensity (-dF/dT) respect to the corresponding temperature, as shown in the figure 13.2. If there is only a single melting peak, it means that no non-specific PCR products are detected in the experiment.
  • Page 92 Gentier Real-Time PCR System User can click the < Analysis Setting > icon in Tool Bar and the application software will automatically pop up the melting curve analysis setting window, which consists of Dye and Gene, Sample and PeakArea three tabs. User can set the relevant parameters for melting curve analytical method in the melting curve analysis setting window.

  • Page 93: High Resolution Melting

    Gentier Real-Time PCR System 3.3.4.4 High Resolution Melting The melting temperature T of dsDNA depends on its base sequence, fragment length and GC content. In theory, any base change can cause difference in melting temperature; however, the difference caused by a base change is tiny, usually less than 1 degree. As the resolution of melting curve is not enough, it is required to read the fluorescence signal several times within one degree of temperature change to accurately detect the difference of melting temperature.
  • Page 94 Gentier Real-Time PCR System Interface Introduction and Parameter Description Reminding: the parameter descriptions of Sample Setting, Result Table and Heat Map functional modules are same with those introduced in D-3.3.4.1 Abs Quant. Melting Curve: display the fluorescence intensity against the temperature, as shown in figure D-14.1.
  • Page 95 Gentier Real-Time PCR System Normalized Melting Peak, Diference Plot, Normalized Melting Curve functional modules and the group calling result will change accordingly. Normalized Melting Curve: display the normalized melting curve according to the normalization method and fluorescence normalization settings. For more details, please refer to D-3.3.4.4 High Resolution Melting - High Resolution Melting Analysis Setting.
  • Page 96 Gentier Real-Time PCR System Figure D - 14.3 HRM analysis interface - Difference Plot functional module Normalized Melting Peak: display the first negative derivative of the normalized melting curve, as shwoin in figure 14.4. The melting temperature range of each sample appears as a peak after normalization, and optionally, temperature shift.
  • Page 97 Gentier Real-Time PCR System Figure D- 15. High Resolution Melting Analysis Setting interface - High Resolution Melting tab ▶ Normalization Method: the application software provides two normalization methods: Ratio Method and Exponential Method. The Ratio Method option is selected by default. ▶...

  • Page 98: Genotyping

    Gentier Real-Time PCR System a. Delta T Discrimination: the default Delta T discrimination sensitivity provided by the amplification software is 0.5; reduce this value, the application software will grouping the melting curve at a higher temperature resolution. b. Curve Shape Discrimination: the default curve shape discrimination sensitivity provided by the amplification software of is 0.5;...
  • Page 99 Gentier Real-Time PCR System Figure D-16 Genotyping analysis interface Interface Introduction and Parameter Description Scatter Plot: display the end point fluorescence distribution of two dyes, as shown in figure 16.1. Figure D-16.1 Genotyping analysis interface - Scatter Plot functional module...
  • Page 100 Gentier Real-Time PCR System ▶ Parameter introduction: X axis and Y axis represent the Relative Fluorescence Unit (RFU) of different dyes respectively; Each point represents a sample; different icons represent different genotypes or interpretation results. ▶ Function key introduction: The introduction of < Y-axis Coordinate Adjustment >...
  • Page 101 Gentier Real-Time PCR System Genotyping Analysis Setting User can click the < Analysis Setting > icon in the Tool Bar and the application software will automatically pop up the genotyping analysis setting window, which consists of Dye and Gene, Sample and Genotyping three tabs. User could set the relevant parameters for the current analytical method in the genotyping analysis setting window.

  • Page 102: End Point Fluorescence

    Gentier Real-Time PCR System threshold for the X axis and Y axis of scatter plot. c. Quality Threshold: user can use keys to set the quality threshold of application software algorithm. Increase the quality threshold value can improve the credibility of grouping result. d.
  • Page 103 Gentier Real-Time PCR System Figure D-18. End Point Fluorescence analysis interface Reminding: the parameter descriptions of Amplification Curve, Sample Setting, Result Table, Raw Curve, Raw Fluorescence and Heat Map functional modules are same with those introduced in D-3.3.4.1Abs Quant. Interface Introduction and Parameter Description End Point Fluorescence Scatter Plot: display the scatter plot of fluorescence intensities measured at end the point of amplification plateau phase, as shown in figure 18.1.
  • Page 104 Gentier Real-Time PCR System End Point Fluorescence Analysis Setting User can click the < Analysis Setting > icon in Tool Bar and the application software will automatically pop up the end point fluorescence analysis setting window, which consists of End Point Fluorescence and Gene and Sample two tabs.

  • Page 105: Instrument Software Operation

    Gentier Real-Time PCR System E. Instrument Software Operation The Gentier instrument can run independently from the control computer. User can use the touch screen or external mouse to operate the instrument system software, so as to realize the experiment program setting and other relevant operations. 1.

  • Page 106: Operation Area

    Gentier Real-Time PCR System ▶ System Status: display the current instrument system status. Initializing: the instrument system is in the process of initialization; Ready: the instrument system initialization is completed, ready for running; Running: the experiment is running; Pause: the current running experiment is paused; Error: instrument hardware or software error, cannot execute any operation.

  • Page 107: Experiment File

    Gentier Real-Time PCR System Reminding: when the instrument system is under Running status, the < Experiment > function key will be converted to < Pause Experiment > function key, user can press this function key to pause the current running experiment. ▶...
  • Page 108 Gentier Real-Time PCR System Figure E-2. Instrument software – Experiment File interface Interface Introduction & Parameter Description s Experiment File Displaying Area: display the pre-existed experiment files or file folders within the instrument system, as shown in figure E-2.1. Figure E-2.1 Experiment File tab - Experiment file display area...
  • Page 109 Gentier Real-Time PCR System User can press the icon of a certain experiment file or file folder in the experiment file ▶ display area to open it. User can slide the slider or press icons to view all the experiment files or file ▶...

  • Page 110: Run Setting

    Gentier Real-Time PCR System Reminding: the instrument software will name the new experiment file or file folder with creation date and time by default. Figure E-2.4 Experiment File tab - keyboard ▶ < Rename >: rename the selected experiment file or file folder, this key is inactivated when the instrument system is under Running status.

  • Page 111: Temperature Setting

    Gentier Real-Time PCR System Reminding: user could also select an experiment file, press Run Setting tab to enter the run setting interface to view or edit the experimental settings. Figure E 3. Run Setting interface Temperature Setting sub tab 2.2.1 Temperature Setting The Temperature Setting sub-tab is selected on the run setting interface by default, as shown in figure E-3.
  • Page 112 Gentier Real-Time PCR System Hot Lid: user could press the check box of Hot Lid to decide whether to utilize the hot lid heating function, and press icons to edit the hot lid temperature. Reminding: the hot lid temperature setting range is 40°C ~110°C. Stage: display the stage type of the temperature program.
  • Page 113 Gentier Real-Time PCR System Temperature Program Setting Operation Step 1: Add stage User can press < Add > in the stage setting box and the instrument software will automatically pop up a stage type selection window, as shown in figure E-3.2. Figure E 3.2 Temperature Setting sub tab Stage type selection window Seven predefined stage types are provided by the instrument software for user: ▶...
  • Page 114 Gentier Real-Time PCR System Figure E - 3.3 Cycle edit box ▶ User could set the cycle number in the cycle edit box, press < OK > to confirm the cycle setting or press < Cancel > to cancel the setting. Reminding: the cycle setting range is 1~99.
  • Page 115 Gentier Real-Time PCR System ▶ Four Step Mode options are provided by the instrument software: Standard, Touchdown, Gradient and Long. User can select a certain step mode option by pressing the icon in front of it, and the icon will turn blue. Figure E-3.4 Temperature Setting sub-tab - Step Edit window - Standard step mode 1) The Standard step mode option is selected by default.
  • Page 116 Gentier Real-Time PCR System Reminding: the temperature setting range is 0.0°C ~100.0°C. Reminding: the time setting range is 1s~60min. Reminding: the ramp setting range is 0.1°C/s~6.1°C/s. 2) If the Touchdown step mode option is selected, the step edit window will be shown as figure E-3.5.
  • Page 117 Gentier Real-Time PCR System Figure E-3.5a Step Edit window - Touchdown step parameter setting Initial Temp.: the initial value of annealing temperature change range. Target Temp.: the target value of annealing temperature change range. Delta Temp.: the temperature change(°C) per cycle. Start Cycle: the cycle number after which the temperature change is started.
  • Page 118 Gentier Real-Time PCR System temperature holding time from the initial time to the target time as the cycling proceeds. ▶ User can press the input boxes of Initial Time, Target Time, Delta Time and Start Cycle to edit the corresponding parameters for the current long step, as shown in figure E-3.6a. Figure E-3.6a Step Edit window - Long step parameter setting Initial Time: the initial value of the elongation time change range.
  • Page 119 Gentier Real-Time PCR System Figure E 3.7 Temperature Setting sub tab Step Edit window Gradient step mode ▶ The gradient step mode allows the Gentier sample to adopt different temperatures. After setting the temperature center value and the temperature offset value for the current gradient step, the system will automatically calculate the gradient temperatures.
  • Page 120 Gentier Real-Time PCR System Reminding: the temperature center setting range is 35.5°C ~99.5°C. Reminding: the temperature offset setting range is 0.5°C ~20.0°C. Reminding: the other parameter settings are same with the standard step mode. Reminding: the Gentier 96S/96C instrument model cannot support gradient PCR step mode.
  • Page 121 Gentier Real-Time PCR System Reminding: the temperature increment setting range is 0.1°C - 5.0°C. If the current step is the last step of a continuous melting stage, the step setting window of this step is as shown in figure E-3.9. Figure E-3.9 Step Edit window - Continuous Melting step mode ▶...

  • Page 122: Fluorescence Setting

    User can set the fluorescence channel and dyes for the current experiment in the Fluorescence Setting sub-tab. User can check the corresponding check box in the Channel column to set the fluorescence channel for the current experiment (1-6 channels for Gentier 96E/96S and 1-4 channels for Gentier 96R/96C); User can press the corresponding icon in the Dye column and select the proper dye from the drop-down list;...

  • Page 123: Run Monitoring

    Gentier Real-Time PCR System 2.3 Run Monitoring After finishing the experiment setting, user can press the < Run Experiment > function key to start running the current experiment, the system will enter the run monitoring interface, which is consisted of three parts: run status bar, run monitoring option bar and run monitoring diagram, as shown in figure E-5.
  • Page 124 Gentier Real-Time PCR System Run Monitoring Diagram: If user select the Fluorescence option in the run monitoring option bar, user can press ▶ keys below the run monitoring diagram to choose the monitor content, which can be the real-time Amlificaiton curve, Melting curve or fluorescence Heat Map of the current experiment, as shown in figure E-5.1.
  • Page 125 Gentier Real-Time PCR System Figure E-5.2 Run Monitoring Diagram - Real-time Melting Curve User can press key below the run monitoring diagram to monitor the fluorescence Heat Map of the current experiment, as shown in figure E-5.3. Figure E-5.3 Run Monitoring Diagram - Heat Map The fluorescence heat map displays the 96 sample wells corresponding to the sample block;...

  • Page 126: General Setting

    Gentier Real-Time PCR System Figure E-5.4 Run Monitoring Diagram - Temperature program 2.4 General Setting The General Setting tab consists of Instrument, Configuration and Service three sub-tabs. 2.4.1 Instrument Sub-tab The General Setting tab displays the Instrument sub-tab by default, as shown in figure E-6.1. Figure E- 6.1 Instument software –...
  • Page 127 Gentier Real-Time PCR System Introduction to Instrument sub-tab ▶ Instrument Serial Number: display the factory serial number of the current Gentier instrument. ▶ Instrument Model: display the current Gentier instrument model. ▶ Instrument Name: display the name of the current Gentier instrument, user can press < Edit >...

  • Page 128: Configuration Sub-Tab

    Gentier Real-Time PCR System 2.4.2 Configuration Sub-tab Figure E- 6.2 General Setting tab - Configuration sub-tab Introduction to Configuration Sub-tab ▶ Network Information: display the network information of the current Gentier instrument. User can press < Edit > and the instrument software will pop up the network information window, as shown in figure E-6.2a.
  • Page 129 Gentier Real-Time PCR System In the IP Setting box, user can view all the network IP addresses, or press < Edit > to modify the corresponding parameter. User can press < Default > to recover the default network IP settings of the current instrument.

  • Page 130: Service Sub-Tab

    Gentier Real-Time PCR System User can press < Confirm > to confirm the date and time setting; User can press < Back > to return to the Instrument sub-tab interface. ▶ Language Setting: user could set the instrument system language. The two options provided are English and Simplified Chinese.
  • Page 131 Gentier Real-Time PCR System ▶ DebugLog: user could connect an USB drive to the Gentier instrument and press press < Get DebugLog > to download all the log file of the current Gentier instrument, this will help our professional engineer to get the relative information in case the instrument require maintenance.

  • Page 132: Instrument Cleaning And Maintenance

    Gentier Real-Time PCR System F. Instrument Cleaning and Maintenance Under proper use condition, the Gentier instrument requires little maintenance. However, the Gentier instrument should be cleaned and maintained on a regular schedule for long time and constant use. This section includes the information on cleaning and maintenance of the Gentier instrument.

  • Page 133: Clean Touch Screen

    Gentier Real-Time PCR System Prohibit: do not spray the detergent directly on the instrument, as malfunctions of the electronics or may occur. Caution: please do not use organic or strong detergent to clean the instrument shell, which may ruin the surface coating. 1.2 Clean Touch Screen step: Switch offthe instrument and unplug the power cord;...

  • Page 134: Instrument Maintenance Operation Instructions

    Gentier Real-Time PCR System 2. Instrument Maintenance Operation Instructions 2.1 Maintain Sufficient Air Flow The placing area of Gentier instrument should be checked regularly, as it requires sufficient air flow to precisely reach the correct target temperature. Please ensure that the air flow is unrestricted and other items around the instrument do not interfering with the air flow.

  • Page 135: Replace Fuse Tube

    Gentier Real-Time PCR System 2.4 Replace Fuse Tube The fuse tube (type F10AH250V) of Gentier instrument is located in the fuse tube box near the power outlet at rear of the instrument. Before replacing the fuse tube, please switch off the instrument and unplug the power plug, use a screwdriver to pry open the fuse tube box.

  • Page 136: Instrument Disinfection

    Gentier Real-Time PCR System Figure F 2. Replace the dust filter foam 3. Transportation or Return to Factory 3.1 Instrument Disinfection In case the Gentier instrument will be moved to another lab or needed to return to the after-sale service department for maintenance, please first disinfection the instrument and fill in the disinfection certificate.

  • Page 137: Install Transport Lock

    Gentier Real-Time PCR System 3.2 Install Transport Lock In order to protect the Gentier instrument during transportation, avoid the transport damages. Please follow the instructions listed below to install the transport lock: S tep: please use appropriate tool to pry off and remove the removable air vent cover located on the right side of the instrument shell, as shown in figure F-3a.

  • Page 138: Product Packaging

    Gentier Real-Time PCR System 3.3 Product Packaging Please use the original packaging materials to properly pack the Gentier instrument and its accessories in order to prevent the collision and oscillation during transportation. Caution: The original transport package of Gentier instrument is designed to reduce the instrument damage and ensure its transportation safety.
  • Page 139 Gentier Real-Time PCR System Figure F - 4. The packing steps of Gentier instrument...

  • Page 140: Roubleshooting

    Gentier Real-Time PCR System 4. Troubleshooting In general, corrective instructions will be display along with the error messages by the application or instrument software. And under normal circumstances the software running errors can be solved by restarting the computer or the instrument system. This section describes the main possible errors of the Gentier Instrument together with possible causes and corrective instructions.
  • Page 141 Gentier Real-Time PCR System Abnormal ramp Air vent is blocked Clean up the air vent. or incorrect temperature Connecting lines are loose. Contact us Caution: in case you cannot judge and eliminate these failures by yourself, please directly contact with our company. Caution: in case any of following situation occurs, please immediately cut off the power supply and contract us.

  • Page 142: Contact Information

    Gentier Real-Time PCR System G. Contact Information Drawell International Technology Limited Chongqing Drawell Instrument CO,.Ltd Add:Suite 2705,Building No.12,Shiyou Road No.1,Yuzhong District, Chongqing,China Tel: 0086-023-63268643 Shanghai Drawell Scientific Instrument Co.,Ltd Add : Suite 1117,Lane561 XiuChuan Rd.,PuDong New Area,Shanghai,China Web : www.drawell.com.cn Email : sales05@drawell.com.cn...
  • Page 143 Drawell International Technology Limited Chongqing Drawell Instrument CO,.Ltd Add:Suite 2705,Building No.12,Shiyou Road No.1,Yuzhong District, Chongqing,China Tel: 0086-023-63268643 Shanghai Drawell Scientific Instrument Co.,Ltd Add : Suite 1117,Lane561 XiuChuan Rd.,PuDong New Area,Shanghai,China Web : www.drawell.com.cn Email : sales05@drawell.com.cn...

This manual is also suitable for:

Gentier 96sGentier 96rGentier 96c

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