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Old 10-13-2016, 10:31 PM   #5
Sylv
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Location: Norway

Join Date: Oct 2016
Posts: 6
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Quote:
Originally Posted by adam.geber View Post
What volumes are you working with? Diluting your sample may help with clumping produced by BSA or other components of your digest.

How long are you drying your beads for? Have you tried doing additional ethanol washes? If you're vortexing, are you then spinning down your elution volume before placing it on the magnet to prevent droplets from coating the sides of your wells?

As jdk787 said, though, this is only a problem if you have reduced yield or bead carryover.
We start out with 500ng DNA in 35Ál, and add 15Ál restriction mastermix so a total of 50Ál. 75Ál beads are then used for purification.
The beads are dried for 5 min. Illumina protocol says at least 15 min, but distributer of the beads says that is way to long.
I don't vortex, but use a shaker for mixing. That has been enough before, but this time I basically had to scrape the beads off with the pipette tips.

BSA in the cutting reaction?...Thanks for making me aware of that.

Last edited by Sylv; 10-13-2016 at 10:49 PM.
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