PCR product purification

well shabeena you must start by runing it on a gel to see how well the pcr was done. Then must purify your pcr product and the best and easy method for me is by using Exonuclase I and Shrimp alkaline phosphatase to remove the excess od primers and other materials not needed. After ur purification step the PCR product should be ready for sequencing which is quite easy from now...
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Hi
I have pcr products, i want to do sequencing,
please suggest how i can proceed for my further work.
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I am trying to run PCR-RFLP and i have purified the PCR product before digesting it. My question is how much PCR product should i use for digestion? The size of the PCR product is 633bp and the enzymes i will be using are HinfI, HhaI, HindIII, MaeII and HaeIII.
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je voudrais avoir un protocol de purification des produits PCR à partir du gel d'agarose. il s'agit de fragment de taille de 650 à 1100pb
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Hi there,
If you're considering using your PCR product as starting material in a restriction digest, you just need to make sure that the final buffer composition of the digest is appropriate for the restriction enzymes you're using. If you're using a large volumem of PCR product in your digest, check the composition of the PCR buffer. Do you need to add less Tris or other salts to make up for the salt already present?
The gold standard is to try the digest without purifying your PCR product, and compare that insert's ability to form colonies after ligation with a purified insert's ability to form colonies. If there's not much of a difference and you get positive colonies even when you skip the purification, then your purification wasn't doing much to begin with.
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