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Primer design

lipase gene pseudomonas sp

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#1 LTSAL

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Posted 04 February 2015 - 08:39 AM

Hi all;

 

Im new in the primer design section, can you please guide on how to design the primer. I need to design a primer to hit my lipase gene. I have the 16S rDNA sequence result.

 

A step by step protocol is much appreciated

 

Regards



#2 memari

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Posted 18 February 2015 - 08:16 AM

I usually use www.origene.com that gives me validated qPCR primers for Human and Mouse. Goto www.origene.com and search for your gene, then choose  primers section for Human or Mouse.

Then Click on Register to view the sequences and put an email address and it gives you the sequences of primers like this one:

 

Human

http://www.origene.c...r/HP220445.aspx

Mouse

http://www.origene.c...r/MP212372.aspx

 

 

or you can go to PRIMER-Blast that has capability to design primer and also to check primers.

http://www.ncbi.nlm....s/primer-blast/

and put your sequence inside PCR Template.

then in Exon/intron selction you can ask to have span exon-exon junction or not.

Choose your animal and cell in Primer Pair Specificity Checking Parameters and also choose mRNA or Total RNA or Genome as database to search and alignment.

in  Advanced parameters you can change the length of primer

Then click on Get Primers button,

 

Babak


Edited by memari, 18 February 2015 - 08:28 AM.

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Babak Memari




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