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Hello,
I have been using the recommended input of 3ug for Aglient SureSelectXT but when I QC post covaris/post clean-up, I am left with only 1-1.5ug of material for the library prep. I've checked my starting ng/ul (using Qubit and tapestation) and have run the samples on a gel, everything looks fine and they are not degraded so I can't understand why I start with 3ug and am left with so little for the library prep.
Does anyone have any experience using 1ug vs. 2ug vs. 3ug for this prep and info on dup rates etc? I was thinking I could use ~5 or 6ug for the covaris and then normalise down to 3ug post clean up just to make sure i'm putting in at least 2ug for the prep.
Thank you!
I have been using the recommended input of 3ug for Aglient SureSelectXT but when I QC post covaris/post clean-up, I am left with only 1-1.5ug of material for the library prep. I've checked my starting ng/ul (using Qubit and tapestation) and have run the samples on a gel, everything looks fine and they are not degraded so I can't understand why I start with 3ug and am left with so little for the library prep.
Does anyone have any experience using 1ug vs. 2ug vs. 3ug for this prep and info on dup rates etc? I was thinking I could use ~5 or 6ug for the covaris and then normalise down to 3ug post clean up just to make sure i'm putting in at least 2ug for the prep.
Thank you!