PCR and RFLP

Elizabeth Osterndorff ElizabethO at mail.utexas.edu
Fri Oct 6 13:06:15 EST 2000


Sheila,

Depending on the sizes of the digested pcr products (if they're not too small), you could also use a high resolution agarose like metaphor from FMC (no affiliation).  It's expensive, but it has worked for me in the past.  I used it for an rflp polymorphism
analysis of a 107 bp pcr product.  The digested fragments were 93 and 14 bp (allele A) and 87 and 20 bp (allele B).  Analysis of the 93 and 87 bp fragments is all that is necessary to determine allelotype, and the high resolution agarose enabled me to
resolve these two fragments. The agarose is a bit of a pain to work with - if you decide you want to try it I can send you some tips that helped me use it successfully.

Good luck!

Elizabeth

"Ma. Sheila M de Jesus" wrote:

> I seek professional advice on this:
>
> I am working on comparing patterns or polymorphism within hemophiliac patients using RFLP on PCR products.  The pcr product from intron 13 generates a 96 bp product.  Is the protocol logical?
>
> Our lab is not capable of doing southern blotting but is capable of doing PCR . This is my research fellowship project and unfortunately, no one here can provide me advice.  Quite a neophyte in the aspect of molecular biology but very familiar with PCR.
>
> If you can share an RFLP protocol in the analysis of the porlymorphism is a big help.
>
> Again, many thanks in advance.
>
> Sheila
> mshmdeje at ustcc.ust.edu.ph
>
> ---
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