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!
Photo
- - - - -

PCR Primer Dilution from F+R


  • Please log in to reply
1 reply to this topic

#1 training_in_molecbio

training_in_molecbio

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 02 December 2010 - 08:17 AM

Ok, I have a question involving making a primer working stock by combining the F+R primer

So, I have my F and R primers seperately at 100micromolar. I want to make 200microliters of a 7.5micromolar working stock. When I set up C1V1=C2V2, I get: V1= [(7.5uM)(200ul)]/(100uM)= 15ul. However, does this 15ul actually mean 30ul since the F and R are seperately at 100uM? To perhaps clarify better, am I adding 7.5ul of the F and 7.5 of the R to 185ul of H20? Or, am I adding 15ul of Forward + 15 ul of Reverse to 170ul H20?

I would greatly appreciate any help.

#2 ivanbio

ivanbio

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 116 posts
7
Neutral

Posted 02 December 2010 - 08:35 AM

Your second option is correct. If you want to have both primers together in one solution at 7.5 uM, you need to add 15 ul of each and then make up the difference with 170 ul of water.

Ivan
Carlsbad, CA




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

©1999-2013 Protocol Online, All rights reserved.