Hi everyone. I have a simple question, but i can't seem to solve it myself. Ive found 2 primer sequences in an article in order to amplify my region of interest. However, I can't find out how to calculate the expected product size. As I'm using a Taqman assay, the product size shouldn't be too big. Can someone help me and explain me step by step how to calculate this?
These are the sequences:
f 5 ′ -ctctgcgctggcaatacag-3 ′
r 5′ -aaatgcaacctaaggag
gagagttc-3′
Hi,
With the known primers for a gene of interest..it should be straight forward..what is the difficulty you are facing exactly is not clear...You can use any sequence analysis program or can get help from some e-PCR application to figure out the exact amplicon size...As you are planning to use a Taqman assay...make sure the available primers amplify the product suitable for the assay....or you may need to design one or both new primers for the assay..
bye
Dear Suri,
I tried searching the homologous sequences in the GenBank. But your primers are showing match to a number of different organisms including human MCM6 gene, drosophila, Equus etc. If you can share some more information on the target genome, I may retry finding the amplicon size. else you can also do BLAST search and get the approximate sixe of amplicon.
Best wishes,
SD
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