PCR-RFLP: precipitate or dilute PCR buffer
Wed Nov 30 20:54:23 EST 1994
I am considering doing some PCR-RFLP. I have a few questions before I
decide what method to use:
1. Are people generally 'removing' the PCR buffer by spin columns or ETOH
precipitation OR simply diluting the PCR buffer before restricting.
2. If changing buffers by spin column or other method, what product/method
seems to give the best cost/performance ration.
3. If diluting have people found some particular enzymes work well and
some not (i.e., they are sensitive to the residual KCl) (I intend to use
4-cutters). What dilution factors are required to give good performance.
4. I am considering visulization via end labelling with P, but
appreciate other suggestions.
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