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
- - - - -

polymerase to use for cloning

cloning polymerase

  • Please log in to reply
4 replies to this topic

#1 srpres

srpres

    member

  • Active Members
  • Pip
  • 17 posts
0
Neutral

Posted 06 March 2013 - 07:31 AM

Lets say I wanted to clone a region of DNA (a 2.0 kB putative promoter region) from genomic DNA into a reporter vector, like pGL3 basic, by PCR-mediated amplification.

Should I use a high fidelity polymerase for the PCR or just regular Taq?

If I use a high fidelity polymerase, which one should I use?

#2 GNANA

GNANA

    GNANA

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 209 posts
9
Neutral

Posted 06 March 2013 - 09:40 AM

always better to go with high fidelity pol for cloning. there are lot available in the market, like Phusion (Neb), platinum pfx , Accuprime Pfx (invitrogen), Pfuultra (stratagene) etc ....i would go with Accuprime, as i am personally used to that.
I would prefer being perfectionist rather than a passionist in Research.

I always had an alternate hypothesis....

#3 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,830 posts
414
Excellent

Posted 06 March 2013 - 11:47 AM

I like KOD, but the pfu range tends to be the standard.

#4 srpres

srpres

    member

  • Active Members
  • Pip
  • 17 posts
0
Neutral

Posted 06 March 2013 - 12:43 PM

I like KOD, but the pfu range tends to be the standard.

What is KOD? What do you mean by "pfu range?" Thanks.

#5 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,830 posts
414
Excellent

Posted 06 March 2013 - 12:56 PM

KOD is KOD polymerase

pfu range means any one of the DNA pols that are derived from pfu (e.g. pfuUltra).




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

©1999-2013 Protocol Online, All rights reserved.