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Primer BLAST

primer blast

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#1 Bassam27

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Posted 31 July 2012 - 04:01 PM

how u know ur primers set that u design is specific to ur target sequences by using blastn ?

and why when amplificating cDNA you need to design primes that cross the exon boundaries ?and if u know any articals that tell the reasen behind that..

what i know is that to be able to distinguish between gDNA produect AND cDNA PRODUCTS .

#2 gyma

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Posted 31 July 2012 - 04:52 PM

1. longer primers provide higher specificity. for example, if you have a primer that has only 3 bases like ATG, then you can imagine its specificity. Generally, a primer length of 25-30 would be enough for a high specificity.

2. you are right. gDNA contains intron so the product would be longer than that from cDNA.





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