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Position: Discussion forums > Bioscience biotechniques discussion forums > Molecular biology troubleshooting forum
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Purification of small (100bp) PCR products
Posted by: monsa (IP Hidden, New member, 9)
Date: March 11, 2005 04:24AM
I am trying to purify a 144bp PCR product. I will use this as a probe template so I want to make sure it is free of other PCR component (i.e., dNTP, taq, etc) and also get rid of an extra, faint band of higher (~300) molecular weight. I tried with the Qiaquick columns (Qiagen) but without success. According to the manual, they are good in the range 100bp - 10 kb, so this may be the problem.
Does anybody have a good purification protocol for small (~100bp) PCR products? Regards, Monsa
Re: Purification of small (100bp) PCR products
Posted by: sheantine (IP Hidden, Unregistered user, )
Date: March 13, 2005 04:20PM
There are several commercial kits and home-made protocols available for purifiy PCR products (100 bp- 10kb). However, they are all not efficient in terms of extreme small or large fragments.
The best experience I have so far is GenCatch gel extraction kit for my 78-bp fragment >60% recovery and >90% purity.
Re: Purification of small (100bp) PCR products
Posted by: Michael L. Altshuler (IP Hidden, Unregistered user, )
Date: March 14, 2005 12:20AM
Try Silica (not the diatoms). It must be good for very short PCR fragments
Re: Purification of small (100bp) PCR products
Posted by: trackie (IP Hidden, New member, 1)
Date: July 7, 2005 06:56PM
You could try Exo-SAP It from Amersham for clean-up, although it will probably not get rid of the higher m.wt band.
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