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Old 06-15-2018, 09:31 AM   #1
aryehhirt
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Default Template Switching RT Basic Q

What exactly happens at the end of template switching RT, and why is it advantageous? Lets assume we are using a MMLV reverse transcriptase to add the ccc at the end of the fragment. What happens next? I could not find a clear explanation online.
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Old 06-15-2018, 05:16 PM   #2
luc
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An DNA oligo with a rGrGrG 3'-end ((the template switching oligo) will anneal to the CCC overhang and the RT will continue to work on this newly added template. This allows you to add a defined (PCR-amplify-able) sequence to the 5'-end of your cDNA.
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