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Summary of Contents for anitoa Maverick qPCR
- Page 1 Anitoa Maverick qPCR Quick Guide For Windows® - based Software Version 1.66...
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Page 2: Table Of Contents
Maverick qPCR Quick Guide Contents Product overview ..........................4 Features .............................. 5 Key parameters ........................... 5 Applications ............................6 Instrument external dimension ......................7 Hardware set-up guide ........................8 Power up the instrument ........................8 Connect to a PC via USB ........................8 Light indicator states ........................... - Page 3 Maverick qPCR Quick Guide Amplification analysis ........................22 Positive vs Negative results ......................24 Standard Curve Analysis ........................25 Steps to perform standard curve analysis: ..................26 Save experiment and Print Report ....................27 Troubleshooting guide ........................28 Issue with connecting the power ....................... 28 The test tube would not fit, or the lid would not pressure on test tube when closed ......
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Page 4: Product Overview
Maverick qPCR Quick Guide 1. Product overview Anitoa Maverick is a portable, high performance real time quantitative PCR system. Maverick is equipped with a 4-channel fluorescence optical system, powered by Anitoa's ultra-low-light CMOS bio-imaging sensor. Maverick is optimally suited for applications where portability, minimal space, fast time-to-result is required. -
Page 5: Features
Maverick qPCR Quick Guide Features Compact and rugged design. No internal moving parts, and no need for calibration. Multi-wavelengths 4-channels fluorescence sensing capability Equipped with ultra-low-light sensitive CMOS-based fluorescent imager. Cross platform software for Windows® and Android® OS, with cloud-ready connectivity. -
Page 6: Applications
Maverick qPCR Quick Guide Temperature control resolution +-0.1 ° C Temperature Ramp Rate 5.5° C/s heating; 4° C/s cooling Size and weight 247mm (L) x 188mm (W) x 133mm (H). Weight 1350g Power supply DC 16V, 90W (Adapter provided that accepts 110V/240V AC) *: Wavelength characteristics can be modified by changing filter modules. -
Page 7: Instrument External Dimension
Maverick qPCR Quick Guide Instrument external dimension Figure 1.2 shows the external dimension of the Maverick qPCR instrument. Figure 1.2 Maverick compact qPCR system product dimension... -
Page 8: Hardware Set-Up Guide
The DC power input connector port is located at the back of the instrument. Once the power is provided, the green power indicator light will turn on. Figure 2.1 Maverick qPCR USB and power connections Connect to a PC via USB The instrument is equipped with an USB port at the back, we can connect the instrument to a Windows®... -
Page 9: Light Indicator States
Maverick qPCR Quick Guide For instruments that are equipped with a touch screen, there is no need to connect the instrument to a host PC or Tablet. Light indicator states There are two LED indicator lights. The green light is used to indicate power supply. As long as the... -
Page 10: Qpcr Tube/Plasticware Requirement
Maverick qPCR Quick Guide qPCR tube/plasticware requirement Maverick qPCR requires the use of full transparent qPCR tubes with flat caps. See figure below for examples. Below are drawings and dimension of the tubes we recommend both in English and Metric units Some examples of the tubes that are proven to work in Maverick instruments: UltraFlux®... - Page 11 Maverick qPCR Quick Guide (https://www.thermofisher.com/order/catalog/product/AB2005) Axygen® 0.2 mL Polypropylene PCR Tube Strips and attached Flat Cap Strips, 8 Tubes/Strip, 8 Flat Caps/Strip, Clear, Product Number PCR-0208-AF-C (https://ecatalog.corning.com/life-sciences/b2b/US/en/Genomics-%26-Molecular-Biology/PCR- Consumables/PCR-Tubes-and-Strip-Tubes/Axygen%C2%AE-PCR-Strip-Tubes/p/PCR-0208-AF-C)
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Page 12: Instrument Cleaning Requirement
Maverick qPCR Quick Guide Instrument cleaning requirement Regular cleaning For everyday cleaning requirement, we can use a soft, clean and damp cloth to wipe the external surface of your instrument. After this step, dry your instrument with another soft cloth. Avoid abrasive cloths, towels, paper towels, and similar items that might cause damage. -
Page 13: Step By Step Software Guide
. It is free. The installed application file will be in C:/Anitoa/Maverick by default. To launch the Maverick software, click the Windows® Start button on the left bottom corner of the desktop and find Maverick software. See below:... - Page 14 Maverick qPCR Quick Guide An icon for Maverick software will also appear on the desktop. We could launch the software by clicking on this icon.
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Page 15: Launching The Software
Maverick qPCR Quick Guide Launching the software When the Maverick PC software is launched, after a brief delay for device initialization, the front page will display as shown in Figure 3.1. Figure 3.1 Maverick software front page Several notable features are marked in Figure 1. Marker (1) is an indicator of whether the Maverick USB device is found. -
Page 16: The Setup Page - Name Setup
Maverick qPCR Quick Guide The setup page – Name setup Click on the “Setup” button will take us to the setup page (Figure 2). Here we can set up the experiment. Here we can name the experiment and all the samples in the wells. We can also name and select the fluorescence channels for qPCR fluorescence reading. -
Page 17: Open Existing Experiment For Template File
Maverick qPCR Quick Guide Open existing experiment for template file We can also load an existing experiment file or template file by clicking on the “Open experiment file” button (Marker (1)). When this step is done, we can click on the “Next step” button to continue setting the thermal cycling program. -
Page 18: The Setup Page - Cycler Setup
Maverick qPCR Quick Guide The setup page – Cycler setup Here we can setup the thermal cycler program. We could choose the type and number of thermal periods and steps, assign time period and temperatures. We could have up to two Pre-Denature periods. -
Page 19: Melting Curve Analysis Period
Maverick qPCR Quick Guide Melting curve analysis period We can also add a Melt Curve (Marker (1)) analysis period at the end of the main thermal cycling program. The Melt curve analysis can be triggered either manually by clicking the start melt analysis button, or automatically by checking “”... -
Page 20: Running The Experiment And Monitor Status
Maverick qPCR Quick Guide Running the experiment and monitor status In the setup page – cycler setup, when we are satisfied with all the settings and loaded the samples, we can start running the experiment by click the “Start…” button. -
Page 21: Running Melt Analysis
Maverick qPCR Quick Guide Running melt analysis When running Melt analysis, the “Run-Melt curve” will show the fluorescence change rate as function of temperature. Figure 3.4. Melt analysis... -
Page 22: Analysis
Maverick qPCR Quick Guide Analysis There are two analysis pages: Amplification analysis and Standard curve analysis. The latter is used for determining the quantity of the DNA targets using standard curve analysis method. Amplification analysis This will show the amplification curves for all samples. We can select the samples and study, set several parameters to this analysis. - Page 23 Maverick qPCR Quick Guide Figure 3.5. Amplification analysis...
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Page 24: Positive Vs Negative Results
Maverick qPCR Quick Guide Positive vs Negative results For each sample’s DNA target, we get a positive vs negative result. Generally, the negative result will be shown as “Neg” and the positive result will be shown with a Ct value between Ct low limit and maximum cycle number. -
Page 25: Standard Curve Analysis
Maverick qPCR Quick Guide Standard Curve Analysis Standard curve analysis is used to determine the absolute concentration of a sample by comparing to other samples with known amount of analyte concentration. We called the samples with known concentration “standards” and the ones with concentration that needs to be determined “unknowns”. -
Page 26: Steps To Perform Standard Curve Analysis
Maverick qPCR Quick Guide Steps to perform standard curve analysis: Step 1: Select the fluorescence channel for such analysis Step 2: Add standard samples – we can select the samples that are standard and click “Add the standard” button (Marker (1)) to add to standard table. We can then assign the concentration numerical values to these standard samples by typing into the table. -
Page 27: Save Experiment And Print Report
Maverick qPCR Quick Guide Save experiment and Print Report Open report page to print report and save experiment. The experiment file will contain all the settings and experiment results. Two report formats will be used for report generation. The excel file report is hand editable. The PDF report is easier to store and use on different platforms. -
Page 28: Troubleshooting Guide
Maverick qPCR Quick Guide 4. Troubleshooting guide Issue with connecting the power Please note that the power connector at the back of the instrument has an orientation. Make sure that the connector is properly aligned with this orientation. Do not force the connector in if the resistance is too great. -
Page 29: The Test Tube Would Not Fit, Or The Lid Would Not Pressure On Test Tube When Closed
Please make sure to follow direction from section “qPCR tube/plasticware requirement” to select the proper test tube. If not sure, just purchase the brands Anitoa recommended or contact Anitoa for samples of the properly sized qPCR tubes. -
Page 30: How Do I Know If The Instrument Is Functioning Properly When Everything Is Plugged In
At this point, we should be able to start an experiment and monitor the reaction well and hot lid temperature change at least. If temperature change does not occur, please contact Anitoa for support of diagnose potential hardware faults. -
Page 31: Touchscreen Blank On Power Up
Maverick qPCR Quick Guide Touchscreen blank on power up Some of the models of Maverick qPCR are equipped with a touchscreen interface. If the touchscreen panel is blank on power up, there are several steps to diagnose this event: 1. Make sure the instrument is indeed powered on. See section “Issue with connecting the power”... -
Page 32: Pcr Data Processing - Positive/Negative Determination And Ct Calculation
Maverick qPCR Quick Guide 5. PCR data processing – Positive/negative determination and Ct calculation. For each reaction well and each channel, a series of cycle-by-cycle fluorescence read out data is obtained. The fluorescence read out from the reaction is processed to determine whether the result is positive or negative. -
Page 33: Additional Data Processing Method - Base Slope Correction
Maverick qPCR Quick Guide fluorescence strength multiplication during the initial exponential growth phase of the positive reaction data. Figure A2 Amplification estimation Additional data processing method – base slope correction Sometimes, when we look at the raw fluorescence data, we see a general negative slope during the initial phase of the data. -
Page 34: Disable Data Processing To View Raw Data For Debugging Purposes
Maverick qPCR Quick Guide Figure A3 Base Slope Correction When we perform curve fitting and calculate the Ct, we would translate and rotate the curve to make the initial phase of the curve aligned to the x-axis of the graph. -
Page 35: False Positive Detection
Maverick qPCR Quick Guide False positive detection The Maverick software uses a set of algorithms to analyze positive reaction data and determines whether positive result determination could be false. When a reaction data is judged false positive, it will show as negative, but with special bracket to indicate that it was due to false positive test. - Page 36 Maverick qPCR Quick Guide (Estimated) Amplification efficiency (value 0 to 100%): The algorithm can estimate the amplification efficiency of the reaction data by calculate the slope of the data when performing sigmoid curve fitting. The optimal amplification efficiency of PCR is 100%, which corresponds to amplicon concentration double every cycle.
- Page 37 Maverick qPCR Quick Guide The user can adjust the threshold values and save them with the experiment file or template file. As shown below, in “Settings”->”Imaging settings” tab the user can selectively adjust the threshold values. Figure A6 Threshold values for false positive determination...
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Page 38: How To Find The Relative Strength, Amplification Efficiency, And Signal To Noise Ratio Of The Reaction Data
Maverick qPCR Quick Guide How to find the Relative strength, Amplification efficiency, and Signal to noise ratio of the reaction data To adjust the thresholds for false positive detection (assuming the user is not satisfied with the default values), it maybe helpful to find out the Relative strength, Amplification efficiency, and Signal to noise ratio of a given reaction data. -
Page 39: Channel Crosstalk Suppression Techniques
The emission spectra of fluorophore materials used in multiplex qPCR assay overlap to some degree. In Anitoa’s Maverick qPCR system, we took several measures to suppress such cross talk both in hardware and software. In this section, we explain the software measures to suppress channel cross talk, using experiment data as illustration. - Page 40 Maverick qPCR Quick Guide In Figure 2 below, we show the result of an experiment. In this example, only channel 2 (HEX) and channel 3 (ROX) is shown. Figure B2. Experiment Data In this experiment, the ROX channel shows strong amplification of concentration ladder products, while the HEX channel gives all negative results.
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Page 41: Method To Adjust Cross Talk Parameters
Maverick qPCR Quick Guide Method to adjust cross talk parameters To illustrate the idea of how cross talk suppression works, let us first go ahead and turn off the crosstalk suppression feature and see what happens. This is done by going to the setting page – step (1), make all crosstalk suppression parameters 0 percent, step (2) let us disable the curve fitting functions to view raw data. - Page 42 Maverick qPCR Quick Guide Figure B4, Cross talk is shown graphically with raw data If we focus on just the hex channel, the effect is clearer: Figure B5, Signal in HEX channel due to crosstalk. As a result, the hex channel signals maybe judged as positive.
- Page 43 Maverick qPCR Quick Guide Figure B6 Cross talk effect in HEX channel is suppressed. So, we can keep 3 percent as the crosstalk suppression ratio for ROX to HEX crosstalk. Now if we turn back on all the curve fitting functions by unchecking the “Curve Fitting Debug Feature”, we will see the curves in Figure 2.
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