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Forum: Sample Prep / Library Generation 12-22-2015, 09:28 AM
Replies: 289
Views: 154,492
Posted By chaichontat
Hi Simone, I'm trying to amplify one gene of...

Hi Simone,

I'm trying to amplify one gene of interest; however, I couldn't get a specific band when I added the whole 10 ul of the RT reaction (final PCR volume was 25 ul). The band only appeared...
Forum: Sample Prep / Library Generation 12-10-2015, 09:18 AM
Replies: 289
Views: 154,492
Posted By chaichontat
Hi Simone, It turned out that my PCR primers...

Hi Simone,

It turned out that my PCR primers were the problem. I was too fixated on the template switching process that I completely ignored the preamplification. Upon replacing the primers,...
Forum: Sample Prep / Library Generation 11-28-2015, 04:40 AM
Replies: 289
Views: 154,492
Posted By chaichontat
Hi Simone, I used the Smart-seq2 protocol...

Hi Simone,

I used the Smart-seq2 protocol using the Superscript IV. For some reason, the reaction failed completely (no cDNA products were observed). When I did not add 1M betaine (Sigma) and 6mM...
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