hercolanium Posted November 2, 2014 Posted November 2, 2014 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’
chadn737 Posted November 10, 2014 Posted November 10, 2014 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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