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primer design


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#1 labster

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Posted 28 April 2009 - 03:17 AM

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!

#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.

#3 labster

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Posted 28 April 2009 - 05:39 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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