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-   -   Smart-seq2 lowest input? (http://seqanswers.com/forums/showthread.php?t=58895)

kobeho24 05-25-2015 11:21 PM

Smart-seq2 lowest input?
 
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!

Simone78 06-04-2015 08:19 AM

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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