Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
- - - - -

qPCR standard for lipase genes

lipase genes standard

  • Please log in to reply
2 replies to this topic




  • Active Members
  • Pip
  • 25 posts

Posted 02 March 2015 - 09:33 AM

Dear all


I have already extracted my RNA and converted them into cDNA. I havent done any cloning into expression plasmids. Is it compulsory to clone the bacteria into these expression plasmids?


How to make standards from cDNA? Can anyone offer some guidelines.



#2 Trof


    Brain on a stick

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 1,300 posts

Posted 02 March 2015 - 02:24 PM

Absolute or relative quantification?


You are asking for standard, so probably absolute, but question is if you really need it.

RNA standards (expressed from plasmids) are rarely used for mRNA quantification, only when no other option exists (i.e. precise quantification of viral RNA), mostly cDNA standards in a linearized vector mixed with some dummy NA. Or most often quantification of genes is done by relatively quantifing it in comparison with control sample.

But if you need the absolute quantity, you need a standard.

Our country has a serious deficiency in lighthouses. I assume the main reason is that we have no sea.

I never trust anything that can't be doubted.

'Normal' is a dryer setting. - Elizabeth Moon




  • Active Members
  • Pip
  • 25 posts

Posted 03 March 2015 - 07:54 AM

Hi currently I'm using a absolute quantification, hence do I need to clone the gene into a vector? Or is it possible to use the purified PCR product and dilute it for standard?

Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.