Nextera half reactions failing after PCR

[michael-steffen]

From what I understand, nextera kits should be able to work at smaller volumes than what the illumina guides say. We have been experimenting with this slightly, and we find that tagmentation works perfecting fine using half reactions(everything scaled to 50% of the normal directions), however, we seem to be losing our samples after PCR. Has anyone else had this problem?

Mike

[dfhdfh]

With "after PCR" do you mean the PCR doesn't work or you lose the samples during PCR clean-up?

[michael-steffen]

Well it could be either/or really. I only checked the samples on the bioanalyzer after the clean up. All full reactions worked, but none of the halfs did.

Mike

[Calluna]

I have not yet tried half reactions, but this post might be helpful. Things you've probably already considered... I would also imagine that accurate pipetting and starting DNA would be very important.