96 well PCR clean up

HMorrison · 07-02-2012, 11:01 AM

Anyone found a 96 well clean up method that will remove 100 nt/bp and smaller from ~250 bp product? Most give 100 bp as the cutoff, but we've gotten too much small material back with Qiaquick 96. Alternative is Ampure, and it can be tricky to handle in 96 well plate. We'd like to clean up, quantitate, THEN pool and size select to get rid of rest of small material before sequencing. Have tried quantitation, pooling, then clean up and size selection, and dataset yields vary too much. Thanks; Hilary

nexgengirl · 07-02-2012, 05:55 PM

We've had success with the 96 Well PureLink Pro kit from Invitrogen

http://products.invitrogen.com/ivgn/product/K310096A

Hope this helps.

ECO · 07-02-2012, 07:32 PM

Nothing will outperform AMPure IMHO. You'll probably have to titrate the cut (specifically the binding time and HOW you mix [static vs constant mixing]) in your own hands, but I would start with 1.3-1.4x for that separation range.

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07-02-2012, 11:01 AM	#1
HMorrison Senior Member Location: Massachusetts Join Date: May 2009 Posts: 116	96 well PCR clean up Anyone found a 96 well clean up method that will remove 100 nt/bp and smaller from ~250 bp product? Most give 100 bp as the cutoff, but we've gotten too much small material back with Qiaquick 96. Alternative is Ampure, and it can be tricky to handle in 96 well plate. We'd like to clean up, quantitate, THEN pool and size select to get rid of rest of small material before sequencing. Have tried quantitation, pooling, then clean up and size selection, and dataset yields vary too much. Thanks; Hilary

07-02-2012, 05:55 PM	#2
nexgengirl Member Location: Maryland Join Date: Apr 2010 Posts: 31	We've had success with the 96 Well PureLink Pro kit from Invitrogen http://products.invitrogen.com/ivgn/product/K310096A Hope this helps.

07-02-2012, 07:32 PM	#3
ECO --Site Admin-- Location: SF Bay Area, CA, USA Join Date: Oct 2007 Posts: 1,358	Nothing will outperform AMPure IMHO. You'll probably have to titrate the cut (specifically the binding time and HOW you mix [static vs constant mixing]) in your own hands, but I would start with 1.3-1.4x for that separation range.