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testing primers efficiency for rt-PCR

primer rt-pcr

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#1 madelingirly

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Posted 27 April 2016 - 08:18 PM

Dear all,

I will design primers for test specific KD of gene in my cell line.

I read on internet, we should test primer efficiency by making STD curve and serial dilution of STD.

so can u give me ur advice?

for example,  if my cell line specifically produced my tested gene, shall I use cDNA prepared from separation of mRNA as STD.

or shall I use other stander, like plasmid or some thing.

Do u have any other recommendation to make good STD curve

Thanks

Shimaa



#2 Andrea Fortina

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Posted 08 June 2016 - 07:24 PM

If you only want to test your primer efficiency, there is no difference in templates, you just need to use same template, and dilute to different concentration, that's will be OK.


Andrea Fortina

Genius only means hard-working for all one's life.

CreativeBiomart, Recombinant protein expert

 

 






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