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a simple primer problem


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5 replies to this topic

#1 zuda

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Posted 02 March 2010 - 05:31 PM

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.

#2 pcrman

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Posted 02 March 2010 - 06:43 PM

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.

#3 zuda

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Posted 02 March 2010 - 06:54 PM

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

#4 Vini

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Posted 02 March 2010 - 08:06 PM

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....

#5 pcrman

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Posted 02 March 2010 - 08:27 PM

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

#6 susanna

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Posted 04 March 2010 - 01:57 AM

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

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






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