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BJS Biotechnologies XXPRESS NGx User Manual

Real-time pcr thermal cycler
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The World's Fastest Real-Time PCR
Thermal Cycler
User Manual
This Manual Copyright © 2018 BJS Biotechnologies Ltd.

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Summary of Contents for BJS Biotechnologies XXPRESS NGx

  • Page 1 The World’s Fastest Real-Time PCR Thermal Cycler User Manual This Manual Copyright © 2018 BJS Biotechnologies Ltd.
  • Page 2 P a g e xxpress, xxplates, xxsealer, and their respective logos are registered trademarks of BJS Biotechnologies Ltd, or one of its affiliated companies in the UK and other countries. All rights reserved. No part of this manual may be reproduced, stored in a retrieval system, or transmitted, by any means or in any form, without the prior permission of BJS Biotechnologies Ltd (BJS Bio).

  • Page 3: Table Of Contents

    P a g e Contents Glossary of Terms and Abbreviations ............. 4 Symbols Used in this Instruction Manual ..........5 Unit Description ..................6 Technical Specification ................9 Safety Precautions and Limitations of Use ..........11 Unit Installation ..................15 Unit Operation ..................

  • Page 4: Glossary Of Terms And Abbreviations

    P a g e 1 Glossary of Terms and Abbreviations ANSI American National Standards Institute BJS Bio BJS Biotechnologies Limited, UK Company Number 02959160 Deepwell Plate Plate with an SBS footprint featuring 48, 96 or 384 wells with a larger volume than microplates...

  • Page 5: Symbols Used In This Instruction Manual

    P a g e 2 Symbols Used in this Instruction Manual The following advisory symbols are used in this manual. Table 1: Advisory Symbol Meanings Indicates a Risk of Electric Shock which could, if not avoided, result in serve injury or death. DANGER Indicates a Burn Hazard which could, if not avoided, result in serve injury or death.

  • Page 6: Unit Description

    P a g e 3 Unit Description The xxpress®NGx Real-Time Thermal Cycler provides a safe and controlled method of heating and cooling samples placed within xxplates™, whilst measuring the level of fluorescence emitted by the samples. xxpress®NGx has five fluorescence measurement channels that enable fluorophores with a wide range of excitation and collection frequencies to be detected.

  • Page 7: Table 2: Unit Features

    P a g e Table 2: Unit Features ® xxpress NGx Power On/Off Button Front Door Front Door Release Button xxplate™ Location ® xxpress NGx Display and Touch Screen Front USB Connector Rear Cover Air Intake Vents and Filters (under unit) Power on LED Emergency Release Button (ECR) Battery Compartment for Emergency...

  • Page 8: Table 3: Xxpress® Ngxsample Plates And Accessories

    P a g e ® The xxpress NGx is designed to be used with a range of sample plates and accessories – some of which are listed below: Table 3: xxpress® NGx Sample Plates and Accessories Picture Description 24 well xxplate™ SBS (sample volume 5-40 µL) 54 well xxplate™...

  • Page 9: Technical Specification

    P a g e 4 Technical Specification Model Type ® Model Name xxpress NGx Thermal Cycler Model Numbers XP-1003-CC1 (220V-240V) XP-1004-CC1 (100V-120V) Physical Unit Properties Dimensions (W x D x H) 300 mm x 520 mm x 315 mm Weight 35 kg Mains Supply Power Cord Rating...
  • Page 10 P a g e | 10 Storage and Transportation -10 to +50 C Temperature Range Relative Humidity Range 20% to 95% non-condensing PCR Performance Thermal Ramp Rate Maximum Ramp Rate 10C per second Adjustable from 0.1C to 10C per second in 0.1C steps Thermal Uniformity ±0.3C well to well across the test plate (Steady state)

  • Page 11: Safety Precautions And Limitations Of Use

    P a g e | 11 5 Safety Precautions and Limitations of Use It is essential that all users of this equipment have fully read and understood the following safety precautions and limitations of use before ® installing or operating the xxpress NGx unit.
  • Page 12 P a g e | 12 Unit Installation and Operating Environment ® The xxpress NGx unit is designed for indoor laboratory use only. The acceptable operating temperature range is 18ºC to 30ºC, DANGER with a relative humidity of 20% to 80% non-condensing, at a maximum altitude of 2000m above sea level.
  • Page 13 P a g e | 13 General Operating Precautions Ensure that the power is switched off at both the AC mains supply outlet and at the back of the unit before inserting or removing the mains power cord. DANGER The xxplate™ can reach temperatures of over 100°C and can remain hot for a time after the xxpress®...
  • Page 14 P a g e | 14 Unit Maintenance and Serviceability There are no user or operator serviceable parts inside the unit. Do not remove the unit casework. DANGER Removal of the unit's casework will void the manufacturer’s warranty and may expose the user to a Risk of Electric Shock resulting in serious injury or death.

  • Page 15: Unit Installation

    P a g e | 15 6 Unit Installation ® Before installing the xxpress NGx unit, please check that the delivery is complete and that the unit and any accessory parts are intact and free from any signs of transportation damage. Also ensure that all external and internal packaging has been removed from the unit before installation.
  • Page 16 P a g e | 16 ® Install and test the xxpress NGx unit using the following procedure: 1) Place the unit on the suitably selected working surface (as specified above), ensuring that the ventilation holes underneath and on the rear of the unit are not obstructed. 2) Open the battery compartment and insert the 9 V battery as shown below.
  • Page 17 P a g e | 17 6) Start-up is complete when the LOGON screen is shown, see below. XXPRESS® NGX USER MANUAL – XP-5001-01-5...

  • Page 18: Unit Operation

    P a g e | 18 7 Unit Operation Please ensure that you have read and fully understood all the Safety Precautions and Limitations of Use listed in Section ® 5 before attempting to operate the xxpress NGx unit. WARNING 7.1 User Touch Screen Display and Controls ®...
  • Page 19 P a g e | 19 • To wake-up the xxpress ® NGx, press the blue Power On/Off Button ® on the front panel once. The xxpress NGx will wake up, run a self- test and initialise its system. Figure 1 is the screen.

  • Page 20: Designing A New Pcr Experiment

    P a g e | 20 Figure 2  Close the screen by touching the symbol in the top right corner of the pop-up window. The following sections of this manual will guide the user through the ® regular operations that can be completed using xxpress NGx.
  • Page 21 P a g e | 21 ® (Choosing the PCR type enables the xxpress NGx to structure the starting thermal cycling profile shown in later screens to match the experimental need). Figure 3 • In this example, we have selected a 3-Step Real-Time PCR. •...
  • Page 22 P a g e | 22 Figure 4 • Touch to select whether based chemistry is PROBE SYBR GREEN ® being used. This information enables the xxpress NGx to preselect the camera sensitivity. Figure 4a • In Figure 4a has been selected. SYBR GREEN •...
  • Page 23 As the xxpress NGx is much faster than traditional thermal cyclers, the BJS Biotechnologies team have developed optimised profiles for running PCR assays at ultra-fast speeds. The figures are only a starting point and all the parameters are adjustable in later screens.
  • Page 24 P a g e | 24 • The next screen controls the thermal profile required for the experiment, see Figure 5. From the information already input into ® xxpress NGx, the system automatically completes this screen with the correct experiment structure and suggested starting values. Figure 5 •...
  • Page 25 P a g e | 25 Figure 5a • Select the parameter to be changed. In figure 5a we have selected the dwell time on step 1 of the PCR cycle. On selecting, the parameter becomes highlighted. Type in the desired value using the keyboard on the right of the display.
  • Page 26 P a g e | 26 up screen. This value can be set from zero to 10°C per second in 0.1°C steps. • The number of times the cycle should be run is set using the REPEAT tile. In Figure 5a the 3-step cycle is repeated forty times. •...
  • Page 27 P a g e | 27 Figure 5b • Touch  the tick or cross to accept or cancel the changes respectively. • At this stage if a Melt Curve is to be run as part of the experiment it can be added as an additional ‘cycle’.
  • Page 28 P a g e | 28 Figure 5d • Touch the tile to add a Melt Curve cycle to the experiment, MELT see Figure 5d. • The Melt cycle is then added to the list of cycles and can be edited by touching.
  • Page 29 P a g e | 29 Figure 6 • Chose the desired xxplate™ SBS size by touching it. The xxpress ® NGx has three xxplate™ sizes: 24 well, 54 well, and 96 well. See Table 3 for the reaction volumes they can accept. •...
  • Page 30 P a g e | 30 • The next set-up screen, seen in figure 7, shows the chosen xxplate™ SBS well layout. • This is an optional page in which the wells can be labelled with information to help with the results analysis. To edit a group of wells, touch to select them, see Figure 7a.
  • Page 31 P a g e | 31 • In the screen that pops up the type of sample that is to be put in the selected wells can be chosen. ▪ refers to the Sample to be tested. SAMPLE ▪ to a No Template Control sample. ▪...
  • Page 32 P a g e | 32 Figure 7d • Touch the tick will accept the information. The process can be repeated as necessary to label other wells on the plate. • You can clear the well selection or even reset the plate completely using the tiles on the right.
  • Page 33 P a g e | 33 • The cell references on the edge of the window make it clear which wells are being viewed, see Figure 7f. Figure 7f • Once all the information is entered as shown in Figure 7f, touch the forward arrow at the bottom right of the screen to move to the review screen.
  • Page 34 P a g e | 34 • Once the user is happy that the experiment is correctly set-up, place the prepared, sealed, centrifuged xxplate™ SBS (see Section 7.2.4) ® into the xxpress NGx. To do this press the Front Door Release Button (See table 2).
  • Page 35 P a g e | 35 • Once the xxplate™SBS is correctly inserted, push the door closed. • Touch the forward arrow on the bottom right of the screen to start the experiment. Figure 9 • Once the experiment is initiated the xxpress ®...
  • Page 36 P a g e | 36 automatically and can now be analysed and/or exported (see Section 7.2.5). Figure 9a XXPRESS® NGX USER MANUAL – XP-5001-01-5...

  • Page 37: Designing A New Melt-Curve Experiment

    P a g e | 37 7.2.3 Designing a New Melt-Curve Experiment • Creating a Melt-Curve Experiment is very similar to creating the PCR Experiment. Select from the PCR menu, see Figure 10. MELT Figure 10 • Touch the arrow at the bottom right corner to advance to the Chemistry screen.
  • Page 38 P a g e | 38 Figure 12 In the Thermal Profile screen, see figure 12, the ‘cycles’ section • shows the temperature range for the melt. The user can adjust this and the number of measurements taken per degree. Figure 12a •...
  • Page 39 P a g e | 39 • Select the type of xxplate™ SBS to be used, see Figure 13. Figure 13 • Advance to the Plate Layout screen using the arrow. Figure 14 • This is an optional page. A plate layout can be created as shown earlier, or alternatively it can be recalled from the memory.
  • Page 40 P a g e | 40 • Touch the desired layout and then accept the selection by touching tick, see Figure 14a. Figure 14a • Advance to the ‘Confirm and Run’ screen using the arrow. Figure 14b • Figure 14b shows the confirmation screen that gives the user the opportunity to review their experiment prior to running it.

  • Page 41: Running The Experiment

    P a g e | 41 • Once the user is happy that the melt is correctly set-up they can insert the prepared xxplate™SBS (see Section 7.2.4). To do this touch the Front Door Release Button (See table 2) to open the loading door.
  • Page 42 P a g e | 42 • To place the prepared and sealed xxplate™SBS into the ® xxpress NGx, touch the Front Door Release Button (See table 2) to open the loading door. Figure 15 • Place the xxplate™SBS onto the slides ensuring that the slides engage into all three holes on either side of the xxplate™SBS, see Figure 15.

  • Page 43: Experiment Results And Analysis

    P a g e | 43 • During operation, it is normal to hear a short hum during each cycle. This is caused by the pressurised air pumps used to cool the xxplate™. • Once the experiment is complete the clamps will release and an information window will pop up to confirm this.
  • Page 44 P a g e | 44 Figure 17 • The Cq is automatically calculated and shown in for each well in the tab, see Figure 17a. The threshold for the Cq ANALYSIS measurements is also calculated and is shown in a blue box on the tab.
  • Page 45 P a g e | 45 Figure 17a • In the melt section, you have the option to differentiate the graph. This helps identify the Tm more easily. Figure 17b shows an example of a Melt in it standard form. Figure 17b •...
  • Page 46 P a g e | 46 Figure 17c • Results can be exported in an RDML or .csv format ready for further analysis. Select the blue tile to do this. Note, if the EXPORT smoothing filter or background filter is active when the data is exported the values will include this filtering.

  • Page 47: Repeating Experiment And Reviewing Previous Results

    P a g e | 47 Note: The RDML export format can be used to directly transfer ® information from xxpress NGx to the BioGazelle PCR analysis program, qBase+. Contact your distributor for more information on qBase+ or go to the BioGazelle web-site, www.biogazelle.com Figure 17e •...
  • Page 48 P a g e | 48 Figure 18 • Then chose the previous experiment that is to be repeated from the pop-up selection box. Touch the to confirm your selection and open the file. The experiment will open at the ‘Confirm and Run’ screen, see Figure 18a.
  • Page 49 P a g e | 49 the experiment by touching the arrow on the bottom right of the screen. • This method of editing and re-running a previous experiment is a quick way of creating new experiments which are similar to ones previously run.

  • Page 50: Maintenance And Servicing

    P a g e | 50 8 Maintenance and Servicing ® Although the xxpress NGx unit does not require any scheduled maintenance or servicing, the operator should regularly clean and inspect the unit for any defects, as described in Section 8.2 below. Please observe and comply with all of the Unit Maintenance and Serviceability precautions listed in Section 8.

  • Page 51: Routine Cleaning And Inspection

    P a g e | 51 Figure 19 1. Remove the AC plug from the power inlet module 2. Use a small flat bladed screwdriver to carefully pull out the fuse holder 3. Fit the correct fuse in the far position only 4.

  • Page 52: Decontamination Procedure

    P a g e | 52 8.3 Decontamination Procedure The unit and accessories should be decontaminated using the following procedure before being stored or transported. Decontamination Procedure Thoroughly clean all outside surfaces of the product (including any accessories, power cords, manuals, packaging, etc) with a damp cloth soaked with suitable disinfectant solution (such as Virkon).
  • Page 53 P a g e | 53 Certificate of Decontamination We respect the health and safety of our clients and employees, and request that any products or accessories being returned are decontaminated in accordance with the procedure below. 1. Decontamination Procedure Thoroughly clean all outside surfaces of the product (including any accessories, power cords, manuals, packaging, etc) with a damp cloth soaked with suitable disinfectant solution (such as Virkon).

  • Page 54: Warranty And Returns

    In no event will BJS Biotechnologies be liable for any indirect, incidental or consequential damages resulting from any defect or warranty claim. Unspecified use or unauthorised modification of any part of the ®...

  • Page 55: Product Disposal

    P a g e | 55 10 Product Disposal At end-of-life, this product must be disposed of in accordance with your local authority regulations for the disposal of potentially hazardous waste and electronic equipment. The unit and its accessories should be decontaminated using the procedure detailed in Section 8.3 before disposal or shipping.
  • Page 57 BJS Biotechnologies Ltd 65 Bideford Avenue Perivale Greenford Middlesex UB6 7PP United Kingdom www.xxpressPCR.com...

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