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Posted

Hello, I have ordered primer set according to the source article, as well as several other articles, should yield a 740 bp PCR products, but when I performed the protocol according to them I get 500 bp PCR products bands when run in gel. When I searched about that I have found another article where they have gotten 600 bp PCR products using the same primer set.

 

The articles with 740 bp:

http://www.apocpcont...emant K Bid.pdf

 

http://biomed.cas.cz...5/zajickova.pdf

 

The article with 600 bp:http://www.lifescien...802_120_131.pdf

 

 

My PCR products and ladder information are the two attached images

 

 

 

What do you suggest I do to make sure if my PCR product is right and how will I know the expected band sizes from the restriction digestion with Taq I?

 

My primers are:

Forward

5-CAGAGCATGGACAGGGAGCAA -3

Reverse

5-CACTTCGAGCACAAGGGGCGTTAGC -3

post-102067-0-33463300-1414923868_thumb.jpg

post-102067-0-86875300-1414923869.jpg

  • 2 weeks later...
Posted

Could be many things. I would sequence some of the products to see what you are getting. Depending on the population or individuals you are working on, you could be seeing natural variation.

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