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
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
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