In my DNA clean up for sequencing and after cleaning amplified DNA with EXoSAP-IT. Is it necessary to wash the PCR product with sephadex? Is there any other clean up kit which may clean up without using sephadex?
I have done a lot of exon sequencing from genomic DNA using PCR amplicons anchored in introns. The only thing I've ever needed is the Qiagen QIAquick PCR Clean up Kit. It's fast and I've never had an issue with it. I simply use the kit, NanoDrop the DNA and send it to our core for sequencing. Done.
there are several quick and cheap kits for PCR clean up. Eric's suggestion is very good, you can see
they are all based on silica columns to which DNA binds or is eluted from depending on the pH of the solutions used.
All companies distributing molecular biology products will offer such products.
Bye, Paola
PCR sequencing:
For me, first I run the PCR product on a gel. Purposes: (1) to check whether my product-of-interest got amplified, (2) whether other non-specific amplicons around. Then, I cut out the unique band and use QIAquick Gel Extraction kit to isolate the DNA out of gel block and send for sequencing. Good results obtained.
QIAquick Gel Extraction kit: https://www.qiagen.com/us/products/catalog/sample-technologies/dna-sample-technologies/dna-cleanup/qiaquick-gel-extraction-kit#productdetails
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