2. Overview In this tutorial we will cover the following topics: Step 1: The wet-lab workflow Step 2: How to select your spike-in during analysis Step 3: Confirmation of sample identity
Hello and welcome to our second interpret tutorial, this tutorial will show you how to use CytoSure Interpret software in conjunction with CytoSure ISCA Sample Tracking Spike-ins.These spike-ins enable you to track your sample through the microarray processing workflow minimising the chance of mixing up your samples.
In this tutorial we will re-cap the experimental process, explaining how the CytoSure™ ISCA Sample Tracking Spike-ins fit into your current workflow.Then we will move on to the data analysis step and show you how to select your CytoSure™ ISCA Sample Tracking Spike-in when using CytoSureInterpret.Finally we will show you how the software checks and confirms the identity of each Spike-in present in your sample.
The CytoSure Genomic DNA Labelling kit with the CytoSure™ ISCA Sample Tracking Spike-ins protocol has been optimised for use when you are setting your labelling reactions up in tube rather than a plate.If you do not have the Spike-in labelling protocol please contact us at products@ogt.co.ukMake sure you follow the protocol carefully, adding one spike-in to each of your samples.As this is the critical step of the protocol we recommend that you get a second person to work with while you set up the reactions so you can be sure of which spike-in has been added to which sample.
After setting up the labelling reaction with the addition of the CytoSure™ ISCA Sample Tracking Spike-ins then the rest of the array processing continues following a ‘normal’ workflow.
Load your data into CytoSure Interpret – if you are not sure how to do please refer to our Quick Start Guide and on-line tutorial which are available from our website. These resources take you, step by step, through the procedure for loading data. The first thing you need to do is tell the software what CytoSure™ ISCA Sample Tracking Spike-in was added to your sampleTo do this click on the edit button within the “Sample information” panel to the right of the screen.
In the dialog box that appears select the ‘other’ tab
In the drop-down list select the correct CytoSure™ ISCA Sample Tracking Spike-in
And then click the update button
If the CytoSure™ ISCA Sample Tracking Spike-in that you have just selected matches the one that is in your sample then the word ‘match’ appears in the “Sample information” panel to the right of the screen.
If they do not match then the words ‘no match’ appear
You can check what CytoSure™ ISCA Sample Tracking Spike-ins are present in your sample by selecting Tools from the menu navigation and then in the drop down list selecting sample identification
In the “sample identification” pop-up window that appears the chromosomes highlighted in orange have a positive match with a CytoSure™ ISCA Sample Tracking Spike-in. To see what spike-in matches that particular chromosome you need to expand this view.To do this you need to deselect the Pool ratios check box.
Now you can see each of the possible spike-ins listed next to the appropriate chromosome. The spike-ins coloured green are those present in your sample.So using the CytoSure™ ISCA Sample Tracking Spike-ins and the spike-in functionality in CytoSure Interpret you can confirm the identity of your sample.
This brings us to the end of the tutorial, please do not hesitate to contact OGT if you have any questions at products@ogt.co.uk
And finally I would like to thank you for your attention.