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Old 11-30-2010, 01:59 AM   #1
mestro2
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Location: germany

Join Date: May 2010
Posts: 4
Default Calibration beads

Hi,

I'm working with the AMPure beads to purify my library preparation products after ligation and PCR steps following the manufactors protocol. But its not possible to get rid of the adapter dimers.Now I hear that you have to calibrate the beads before using it. Is that rigth? And how should I do that?
May be its a naiv question, but I can't find any protocol or help!

Thank you.
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Old 11-30-2010, 07:59 AM   #2
niceday
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Location: cambridge

Join Date: Apr 2010
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Default

Quote:
Originally Posted by mestro2 View Post
Hi,

I'm working with the AMPure beads to purify my library preparation products after ligation and PCR steps following the manufactors protocol. But its not possible to get rid of the adapter dimers.Now I hear that you have to calibrate the beads before using it. Is that rigth? And how should I do that?
May be its a naiv question, but I can't find any protocol or help!

Thank you.
Try two ampure cleanups with 1.8:1. I find with SOLiD libraries the 2nd cleanup really gets the dimers whereas with only one cleanup there is still loads of dimers left.
WRT calibrating the beads we haven't done that.
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