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
- - - - -

Invitrogen platinum pfx enzyme not so fantastic


  • Please log in to reply
1 reply to this topic

#1 heidimb2000

heidimb2000

    member

  • Active Members
  • Pip
  • 6 posts
0
Neutral

Posted 10 August 2004 - 11:21 AM

Hi!

I posted a problem earlier about my pcr product not migrating from the agarose gel well.

I was just informed by invitrogen customer service that:


The problem you have mentioned is one that we have observed before with
Platinum® Pfx DNA Polymerase. Please see the notes below.

Problem: Ethidium bromide-stained DNA in the wells of the agarose gel
following
amplification with Platinum® Pfx DNA Polymerase.
Solution: This is likely caused by a combination of the following:

* Platinum® Pfx DNA Polymerase has more protein per unit of enzyme than
other amplification
enzymes.

* Platinum® Pfx DNA Polymerase may have different DNA-binding
properties
than other
enzymes. If the protein remains bound to the DNA, migration is
decreased in
the gel. If this
reduces the yield of product migrating into the gel, add SDS to the
sample
loading buffer to a final
concentration of 0.1%.

I thought that this might help someone else with this problem. I might suggest using another enzyme if blunt-ended product is needed!

#2 SVTX

SVTX

    member

  • Active Members
  • Pip
  • 28 posts
0
Neutral

Posted 12 August 2004 - 07:22 PM

Hey,

So, it was the protein! I have been quite successful with Stratagene's Pfu. Works great all the time!

SV




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

©1999-2013 Protocol Online, All rights reserved.