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
- - - - -

Taq introduces error after?


  • Please log in to reply
4 replies to this topic

#1 jiajia1987

jiajia1987

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 212 posts
0
Neutral

Posted 17 April 2009 - 06:25 AM

Dear peeps,

I am doing an error-prone PCR and I am interested in introducing mutations in my gene. I have two pairs of primers flanking my gene, which is nearing 2kb in length. Does anyone know on average, Taq introduces a wrong nucleotide during extension after approximately how many base pairs? I have been trying to find this in journals, but I am unable to find any. please help!!!! :blink:!!

Thanks in advance!!

#2 mastermi

mastermi

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 82 posts
0
Neutral

Posted 17 April 2009 - 09:12 AM

I think it's one in a thousand. If you want to produce a lot of point mutations better use additional mutational conditions.

#3 jiajia1987

jiajia1987

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 212 posts
0
Neutral

Posted 21 April 2009 - 05:01 AM

I think it's one in a thousand. If you want to produce a lot of point mutations better use additional mutational conditions.


Thanks for getting back to me. I am sorry for this late reply.

I am using MnCl2 and biased dNTPs to increase the mutations

#4 HomeBrew

HomeBrew

    Veteran

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 930 posts
16
Good

Posted 21 April 2009 - 06:30 AM

Does anyone know on average, Taq introduces a wrong nucleotide during extension after approximately how many base pairs?


This is known in other circles as the "gambler's fallacy". If the rate of misincorporation is 1 in 1000, then *every* base has a 1 in 1000 chance of being wrong, just as there's no better chance of getting a good card hand just because you lost the last six hands in a row.

This assumes Taq is not more likely to misincorporate one base over another.

There are some error rate tables here, but reported error rates vary because they are condition dependent.

#5 jiajia1987

jiajia1987

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 212 posts
0
Neutral

Posted 27 April 2009 - 12:19 AM

Does anyone know on average, Taq introduces a wrong nucleotide during extension after approximately how many base pairs?


This is known in other circles as the "gambler's fallacy". If the rate of misincorporation is 1 in 1000, then *every* base has a 1 in 1000 chance of being wrong, just as there's no better chance of getting a good card hand just because you lost the last six hands in a row.

This assumes Taq is not more likely to misincorporate one base over another.

There are some error rate tables here, but reported error rates vary because they are condition dependent.


Thanks for the error rate table. :D




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

©1999-2013 Protocol Online, All rights reserved.