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Thread | Thread Starter | Forum | Replies | Last Post |
SMART Concatemeres | sylvain | Sample Prep / Library Generation | 4 | 03-09-2018 02:26 AM |
SMART-seq2 and Nextera XT DNA sample preparation kit | wacguy | Sample Prep / Library Generation | 9 | 02-09-2015 01:00 AM |
First index always lowest percent reads | creeves | Illumina/Solexa | 12 | 05-22-2013 06:35 AM |
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#1 |
Member
Location: HKUST, Hong Kong Join Date: Apr 2015
Posts: 32
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Hi All,
I tried to figure out what is the lowest input of Smart-seq2 on our hand. It doesn't work when the input of total RNA amount is 0.1pg or 1pg (serially diluted with RNase Free water), while products with >10pg input looks just fine in bioanalyzer. Have you guys ever tried such assays? Best! |
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#2 |
Senior Member
Location: Bonn (Germany) Join Date: Oct 2010
Posts: 193
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never tried with total RNA below 10 pg but I can tell you that even T or B cells work (and they have VERY little mRNA!), you just have to increase the number of cycles a bit (21-22). In that case, you might have also to decrease the amount of primers and block all the oligos with biotin at the 5´-end, to avoid getting just adaptor dimers.
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