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