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Old 05-22-2017, 01:10 AM   #12
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Location: London

Join Date: May 2017
Posts: 7

Originally Posted by Simone78 View Post
I cut reaction volumes by a factor of 10 using a liquid handling robot and the Nextera kit (exactly the same protocol, no changes). Tagmentation is done in 2 ul, I add 0.5 ul NT (or 0.2% SDS) followed by 1 ul i5/i7 (1:5 dilution to avoid excess adaptors and save money) and 1.5 ul NPM. Input is 50-150 pg/rxn.
Even without a robot this is perfectly doable with a pipette.
I this way a 3000 USD kit will last for about 1000 samples instead of 96!

Dear Simone and other Nextera users,

I was wondering how to go about the Tagmented DNA Clean-up in the Nextera protocol. It uses a Zymo spin column, which I guess will not be possible for a smaller volume (e.g. 2-10 ul). What do you do at this stage? Do you clean with beads? Skip the whole clean-up? Dilute the Tagmented DNA in buffer to make up for the original 50ul volume needed?



Would it b
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