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Old 10-05-2015, 11:22 PM   #161
Simone78
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Location: Basel (Switzerland)

Join Date: Oct 2010
Posts: 208
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Quote:
Originally Posted by kobeho24 View Post
Hi Simone,
Glad to know that you have already tried the TGIRT on your hand. I wonder if 0.2% SDS can strip the sticky enzyme off the DNA efficiently. And have you tried SDS along with high temperature incubation and also RNase H/ RNase A treatment? From the bioanalyzer, things seem much better than the retroviral RT. Hope to see its application in single-cell RNA-seq.

Gary
Hi Gary,
I just tried with 0.2% SDS, in the same way as I am doing after tagmention to strip the Tn5 off the DNA before PCR and it didnīt work. I also tried SDS + NaOH/HCl (skipping columns or precipitation, as they recommend in the protocol) but, as I said, there was probably too much salt at that point and the PCR didnīt work.
/Simone
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