Smart-seq2 lowest input?

[kobeho24]

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]

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.