Quote:
Originally Posted by kobeho24
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!