I'm looking to add a SPRI cleaning step into my workflow after the cDNA generation. Right now, I use the Clontech Smartscribe enzyme to generate my cDNA from RNA. I usually just use some portion of the reaction into a 50uL reaction volume for amplification.
I've heard that some people have had success using Ampure beads to clean the cDNA, then either using the elute, or directly amplify by adding the beads to PCR reaction.
I'm wondering is there a different proportion/protocol for using these beads on single strand products compared to double strand products (as they usually are in regular PCR)? I know the old clontech protocol seems to use SPRI beads to clean cDNA, but the new ones don't seem to bother.
I've heard that some people have had success using Ampure beads to clean the cDNA, then either using the elute, or directly amplify by adding the beads to PCR reaction.
I'm wondering is there a different proportion/protocol for using these beads on single strand products compared to double strand products (as they usually are in regular PCR)? I know the old clontech protocol seems to use SPRI beads to clean cDNA, but the new ones don't seem to bother.
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