Volumetric scaling for library prep

[adam.geber]

Hi folks,

Looking over some of the early papers showing the equivalence of volumetrically scaled Nextera reactions has me wondering which commercial kits aren't amenable to this modification. Is it reasonable to assume that if a) library complexity isn't an issue and b) pipetting is accurate then scaling reactions down should preserve the functionality of the reagents? I'm interested in responses to this question for both manual and automatic operations but I understand the additional variables to consider for the latter.

Thanks!

[nucacidhunter]

I would be cautious about scaling down the RNAseq for expression analysis because it introduces another variable. Having said that if all preps for an experiments were scaled down I do not see any issues. Any DNA prep with the condition that you describe can be scaled down and it works fine. Some companies are smart and their workflow includes addition of 1 or 0.5 ul of reagents in some steps which makes it realy difficult for scaling down.