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IT-IS Life Science MyGo Pro, User Manual

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IT-IS Life Science MyGo Pro User Manual Download Page 43

AUTOMATIC HIGH RESOLUTION MELTING 

DIFFERENCE, NORMALIZED AND MELT

   

Once opened the analysis module will automatically determine 

optimised parameters and generate a set of D ifference, 

Normalized  a nd  M elt g raphs.  

CLUSTERING  

The difference curves will then be converted into a clustering 

graph showing clusters of different genotypes. 

GENOTYPES

   

The software will automatically determine I nferred  

genotypes b ased on the clustering graph. You can define your 

control G enotypes u sing the G enotypes p ane. 

Summary of Contents for MyGo Pro

Page 1: ...PERFORMANCE MADE EASY USER GUIDE MyGo...

Page 2: ...convenient format Novel Full Spectrum Optics deliver 120 optical channels of fluorescence data from every tube for exceptional multiplex PCR High performance Peltier elements combined with solid silve...

Page 3: ...om setting it up on your workbench to configuring the instrument and installing the software TIPS Learn some useful tips when using your instrument EXPERIMENT Learn the basics of setting up an experim...

Page 4: ...LE 4 ETHERNET CABLE 5 USB DRIVE Please keep the flight case and outer box the MyGo Pro came in in case you need to protect your MyGo Pro during storage or shipment in future Lid USB Port SETUP INSTALL...

Page 5: ...es display LEDs to help you understand what the instrument is doing Here is a summary of these display LEDs Blue Initialising Green Idle Red Lid open Please close the lid Yellow Tubes Loaded Blue Runn...

Page 6: ...s three connections One in the front and two in back as shown below This is for connecting your MyGo Pro to your LAN or computer This is to connect your MyGo Pro to the provided power supply unit ETHE...

Page 7: ...lab bench and then connect AC power to the power supply unit Your instrument will now turn on after a few seconds The display LEDs will light up blue and then turn green if a lid is closed or red if n...

Page 8: ......

Page 9: ...ot Please do not touch it The heated lid will be preheated to 105 C if user activity is detected This enables your run to start as soon as possible After 5 minutes of inactivity the heated lid will be...

Page 10: ...version of the software can also be downloaded from our website mygopcr com WINDOWS Double click on the Windows installer and follow the on screen instructions to install your MyGo Pro software on Win...

Page 11: ...With your MyGo Pro software open and your MyGo Pro connected to the network please connect to your MyGo Pro Instrument Please select Configuration CONFIGURING YOUR MYGO PRO...

Page 12: ...To add your new MyGo Pro select the Add b utton You will now be presented with a list of available instruments Please select MyGo Pro...

Page 13: ...Double click on the instrument you wish to connect to or pr ess Select with the instrument selected The instrument will now be added to the list of Registered Instruments for use in the software...

Page 14: ...UR LAB CLEAN 6 KEEP YOUR INSTRUMENT CLEAN 7 PC SETTINGS LID GETS WARM The MyGo Pro lid can get warm during operation this is completely normal SPIN YOUR TUBES This will ensure all well contents are at...

Page 15: ...pace clean including all lab equipment like surfaces pipettes and tube racks This will keep the instrument clean and help maintain good results KEEP YOUR INSTRUMENT CLEAN We recommend a routine cleani...

Page 16: ...earn how to create save open and close experiments You will also learn how to set up an experiment including thermal profile sample information and optical settings Finally we will show you how to run...

Page 17: ...be displayed as shown below By default the experiment name will be N ew Experiment with a date and time stamp which can be edited The summary will give you information about the instrument you are ru...

Page 18: ...ype drop down menu gives you the option to open an experiment normally or as a template that can be used for a new run OPEN AN EXPERIMENT By selecting O pen y ou can open an experiment as shown below...

Page 19: ...and analyses to re run a previous experiment generating new data When an experiment file is opened as a template it has the same 1 Temperature profile 2 Optical settings 3 Samples and targets 4 Analy...

Page 20: ...en starting a run you will be prompted with the window displayed below Make sure the first checkbox is selected and your experiments will be automatically saved To prevent the software from showing th...

Page 21: ...ent is saved you will be prompted to select a location and filename to use After this you can click Save to use the same location overwriting the previous version or Save As to choose a new location C...

Page 22: ...sion and optical data acquisition Typically optical data is acquired at the same temperature that strand extension occurs By default this time is set to 30s However this time can be reduced to a minim...

Page 23: ...shown below PROGRAMS To add a program to your experiment click A dd i n the P rograms p ane Programs can be deleted by selecting them in the list then clicking D elete The program order can also be a...

Page 24: ...e Profile p ane in the order they are present in the P rograms window as shown below PROGRAM SETTINGS To alter the settings of a particular program you should first select it then refer to the right h...

Page 25: ...men being analysed e g Mouse 5 A molecular target e g x174 detected with a specified fluorescent reporter e g FAM Samples or targets do not need to be defined before starting a run They can be added t...

Page 26: ...sign wells by selecting them and clicking Set Samples can also be unassigned by selecting wells you wish to un assign and clicking Clear ASSIGNING TARGETS TO WELLS Targets can be assigned to wells as...

Page 27: ...allows you to calibrate new dyes that are not included in the factory calibrated list The calibration is based on a PCR amplification using the relevant dye or labelled probe To generate your own dye...

Page 28: ...you wish to save or open sample and target information setup you can select Save As or Open respectively CSV Editable csv files PSD A proprietary locked file format RDML The Real time PCR Data Markup...

Page 29: ...your optical data will be The optical settings can be separated in these two different categories INTERCALATING DYES These dyes for example SYBR Green I Resolight Eva Green are bright reagents and re...

Page 30: ...the current dyes present in your experiment You have the following options when working with dyes in your MyGo Pro software OPEN Open dye files from a file to be added to this experiment SAVE Save a d...

Page 31: ...ing saturated by very bright fluorescent reporters For your convenience at the end of every run your MyGo Pro will record a Recommended Amplification Integration Time in the Run Log Runs where every w...

Page 32: ...ed Instruments Select an instrument and press S elect t o begin the run STARTING A RUN FROM A USB DRIVE To start a run using a USB drive select Start Run Then when you are asked to choose an instrumen...

Page 33: ...heated lid is balanced please make sure that the mount contains at least a strip in rows A and D as shown in blue below or single tubes in wells A1 A8 D1 and D8 These positions can be filled with tub...

Page 34: ......

Page 35: ...quantities of template or relative expression levels of genes using Absolute Quantification or R elative Quantification GENOTYPING Use TaqMan probes or High Melting Resolution to perform SNP genotypin...

Page 36: ...with minimal user intervention Manual analysis types enable users to apply traditional analytical methods SELECT ANALYSIS TYPE Once your experiment has completed you can add an analysis type by selec...

Page 37: ...Performance is a measure related to the observed efficiency of amplification at later cycles This value will tend to be lower for inhibited or out competed reactions and higher for efficient reactions...

Page 38: ...Graph Axes drop down menu All 3 thresholds are used for calling even if they are not currently displayed on the graph SETTINGS The software uses a combination of Quality Q Performance P and Intensity...

Page 39: ...exclude this number of cycles from data before performing analysis This feature can be disabled by deselecting this option ARTEFACT FILTERING Biochemical and physical factors can cause fluorescence l...

Page 40: ...None Background Full Background Correction Normalisation BACKGROUND CORRECTION Background fluorescence including drift is estimated and removed There are many potential sources of background fluoresc...

Page 41: ...ifiers Here you will be shown the relationship between Cq and input template quantity plus a variety of statistics derived from the standards RESULTS AS TABLE The results of amplification analysis are...

Page 42: ...Genotyping Genotypes of samples are determined by the ratio of endpoint fluorescence between two TaqMan probes Thresholds for the fluorescence ratios are generated automatically as shown below A utoma...

Page 43: ...f D ifference Normalized a nd M elt g raphs CLUSTERING The difference curves will then be converted into a clustering graph showing clusters of different genotypes GENOTYPES The software will automati...

Page 44: ...MELTING AUTOMATIC TM CALLING Add an Automatic Tm Calling analyses to identify and characterise melting peaks You can see the results below...

Page 45: ...ines how big a peak should be before it is called Increasing this value will increase specificity of peak detection Decreasing this value will increase the sensitivity of peak detection PEAKS AND MELT...

Page 46: ...alysis To fuse multiple experiments together hold down cmd on a Mac or Ctrl on a PC and select the files you wish to fuse from the open dialog window as shown below Select Open and then from the next...

Page 47: ...tiple file locations you must select the first experiment and then select the from the top left of the open file dialogue window as shown below Now hold cmd Ctrl and select the next experiment Select...

Page 48: ...inished analysing your data in the MyGo software you can export the results in a variety of ways from raw data to user defined custom reports You can export data from panes with results in tabular for...

Page 49: ...edited RDML The software is able to export Cq values and amplification curves from Auto Quantification and melt curves from Automatic Tm Calling in RDML format T o do this select S ave as as if you w...

Page 50: ...able CSV or Portable Document Format PDF PNG This is a bitmap image SVG A Scalable Vector Graphic is an XML vector based graphic that can be used where a high resolution scalable graph is required for...

Page 51: ...under the Experiment tab This provides you with the ability to choose which parts of the data to include in a report For example items such as thermal profiles sample information and various analysis...

Page 52: ...MAINTENANCE This section will help you take care of your MyGo Pro It covers the following areas 1 CLEANING 2 DISPOSABLES 3 ENVIRONMENTAL CONDITIONS 4 DISASSEMBLY...

Page 53: ...com Please note that for health and safety reasons you must print and complete a physical copy of the decontamination form and include this with any instrument or lid that is returned The form must b...

Page 54: ...roducts C Disposables should have reproducible wall thicknesses which are thin enough to ensure rapid temperature equilibration but thick enough to avoid breakages In many cases third party disposable...

Page 55: ...of the instrument including the heated lid and the vents on the instrument base and back ENVIRONMENTAL OPERATING CONDITIONS DISASSEMBLY ENVIRONMENTAL STORING TRANSPORTING PACKING CONDITIONS Humidity...

Page 56: ...ered on overnight All experiments can be used as a template Click Open and then using the file type selection in the file dialog to select the Template option then select the file to open as a templat...

Page 57: ...ased Auto Quant fits a model of a PCR amplification to the fluorescence data observed This model fit then enables the estimation of a number of important parameters including Cq values Correct backgro...

Page 58: ...r MyGo Pro will let you know when something is wrong by displaying an error Most errors are reported as messages in the Status Bar in the MyGo software Some errors are reported by the instrument displ...

Page 59: ...ou may need to contact technical support with the following actions described below Saturation Warning Run Prepare Error followed by one of the following messages a No tubes loaded b Instrument has ha...

Page 60: ...code Please contact Technical Support May be reported at the start of a run Please check that your network is properly configured and is operating at 100Mb s or more Please check that your network is...

Page 61: ...HARDWARE ERRORS For Hardware faults go to Configuration MyGo Pro and select Retrieve Instrument Report Files ERROR LOG FILES In the unlikely event that you encounter a problem the software will log t...

Page 62: ...NOTES...

Page 63: ...All trademarks are the property of their respective owners Design and specifications are subject to change without notice...

Page 64: ...www mygopcr com Version 001 MyGo...

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