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Old 02-17-2014, 06:04 PM   #1
wacguy
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Location: NC

Join Date: Sep 2012
Posts: 24
Default SMART-seq2 and Nextera XT DNA sample preparation kit

Hi,

I was preparing 3 libraries for RNA-seq starting with 0.03, 0.3 and 3ng total RNA using the modified protocol for SMARTer ultra low (Picelli et al, 2014). I started the Tagmentaion/amplification step using the Nextera XT DNA sample prep kit with 1ng in each library. It looks to me very promising considering the low input but I think that there is still a place for improvement (see attached file). My impression, and from what I read, is that my DNA input for the Nextra protocol (1ng) is too high. I'm planning to try 3, 4 dilutions to see if I can get a smaller average fragment size and more "narrow" distribution but'd be happy for ideas and suggestions.

Thanks,
Guy
Attached Files
File Type: pdf 140214-111113-20half-roots_SMARTer2_10_final_PCR.pdf (1.19 MB, 300 views)
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