I need to detect the expression levels of two family members of a gene. They are very similar in sequences. How to design primers to distinguish them? In other words, how do I make sure one primer doesn't amplify both members?
many thanks!
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#2
phage434
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Posted 28 April 2009 - 04:32 AM
A 3' mismatch almost assures that a primer will not extend. If the primers are located near a region in which the sequences diverge, this can be used to distinguish the sequences.
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labster
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Posted 28 April 2009 - 05:39 AM
phage434, on Apr 28 2009, 04:32 AM, said:
A 3' mismatch almost assures that a primer will not extend. If the primers are located near a region in which the sequences diverge, this can be used to distinguish the sequences.
