10 October 2014 5 2K Report

Good Afternoon Everyone

I am planning on purifying my PCR product and running three separate restriction digests on that product. The enzymes are BstUI (CG/CG), TaqI(T/CGA), HpyCH4IV(A/CGT).

I have used the Qiagen gel extraction kits before for PCR clean-up...a) the product recovery isn't the greatest b) the process is relatively time-consuming.

I recently came across the Affymetrix ExoSAP-IT For PCR Product Cleanup kit that uses Exonuclease I and Shrimp Alkaline Phosphatase (SAP) to remove single-stranded primers/DNA products and dNTPs. The reaction from start to finish is 30 minutes (15 minutes incubation, 15 minutes heat-inactivation) with no loss of PCR product. 

Would you recommend using this procedure over the gel-extraction one ? I was wondering if there would be any interference with the subsequent restriction digests even after heat-inactivation. 

Any advice would be very much appreciated.

Ani

PS. Has anyone tried the Wizard gel-purification kit from Promega ? Is it any better than the Qiagen one ?

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