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Old 05-26-2015, 12:48 PM   #2
Simone78
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Location: Basel (Switzerland)

Join Date: Oct 2010
Posts: 207
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Quote:
Originally Posted by kobeho24 View Post
Hi all,
Recently, I was wondering that is it necessary to perform a purification step after cDNA generation and prior to library construction? Because I am gonna do the cDNA generation in a very tiny volume. The library construction can be either ligation-based or tagmentation-based.
Best!
Gary
itīs not necessary at all! Just dilute the PCR product to reduce the amount of salt, take an aliquot for ligation/tagmentation and go on. I tried it myself and it works quite well. I never compared the efficiency with a "standard" protocol, though. Based on what I see on the Bioanalyzer (havenīt sequenced any sample yet) I would be surprised to find out that the difference is so dramatic.
good luck!
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