a simple primer problem - (Mar/02/2010 )

hi i have a question, suppose i make a set of primers- one forward and one reverse.. on making the primer when i do blast i find that one primer- say left primer is binding only at one locus in homo sapies but the other primer is binding at different chromosomes, so will the overall product size vary, or not.

-zuda-

most likely you won't get any product if the two primers are not on the same chromosome or are far away from each other.

-pcrman-

NO THAT IS NOT THE CASE, THE PRIMERS ARE ON THE SAME CHROMOSOME , BUT ALSO ONE OF THEM IS FOUND AT ANOTHER CHROMOSOME

-zuda-

zuda on Mar 3 2010, 09:24 AM said:

NO THAT IS NOT THE CASE, THE PRIMERS ARE ON THE SAME CHROMOSOME , BUT ALSO ONE OF THEM IS FOUND AT ANOTHER CHROMOSOME

in that case, u will get the product with primers on the same chromosome....not the other one....

-DRN-

I agree. the mismatch won't be a problem as long as the other does not has mismatch.

-pcrman-

you only have amplication, when both primers bind your product. The same thing using an extra probe to increase specificity

pcrman on Mar 2 2010, 09:27 PM said:

I agree. the mismatch won't be a problem as long as the other does not has mismatch.

-susanna-