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Has anyone used AMPureXP for ssDNA purification? I need to get rid of smaller cDNAs (<100 ntd).
Biochembug
Has anyone used AMPureXP for ssDNA purification? I need to get rid of smaller cDNAs (<100 ntd).
Biochembug
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Yes it works for ssDNA. It also works for RNA. And DNA/RNA hybrids.--------------
Ethan -
What volume should I use to eliminate low molecular weight (<100 ntd) cDNA?Comment
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What I do is I mix 1 volume of DNA or RNA with 1 volume of AMPure XP and 1 volume of 30% PEG800 1.25M NaCl. That gives me a cut off of about 100 bp. If you search the forum there is a thread with more information on this with some guy that posted a nice experiment. Be warned that there is batch to batch variation with the AMPure XP beads.
There are some protocols on my blog at http://ethanomics.wordpress.com/ which perhaps describe what I do a little better.--------------
EthanComment
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Thanks for the info Ethan. Will give a shot!
BiochembugComment
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Hi Ethan,Originally posted by ETHANol View Post
Can you comment on what the cutoff seems to be for dsDNA vs ssDNA? Does it appear to be a function of molecular weight, strand length, or something else?
PhillipComment
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There's a recent post on James Hadfield's blog detailing how the beads work and how the concentration of PEG/NaCl effects the size selection.
Comment
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Hi all,
Great to see that you guys are keeping this thread alive. As pointed out by Phillip, I am looking for the cut-off of the single stranded DNA molecules on AMPure beads. Comments are always welcome!!!
Thanks TonyBrooks for that like.
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