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Old 10-14-2019, 02:45 AM   #16
pabanga1
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Location: Freiburg im Breisgau

Join Date: Oct 2019
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Quote:
Originally Posted by lapensee View Post
FWIW I got this working. I adopted the OMNI-ATAC protocol and get great library amplification from 5k cells using only 2% the amount of Tn5 required for 50k cells. Adding a protease inhibitor during transposition was crucial for my library prep.
Hi! I am wondering what does exactly from the Omni-ATAC protocol mean:

"Add 50 ul cold ATAC-Resuspension Buffer (RSB) containing 0.1% NP40, 0.1% Tween-20, and 0.01% Digitonin and pipette up and down 3 times.
Incubate on ice for 3 minutes.
Wash out lysis with 1 ml of cold ATAC-RSB containing 0.1% Tween-20 but NO NP40 or digitonin and invert tube 3 times to mix"

Does this mean I need to removed the previous 50 uL and wash with cold ATAC-RSB?
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