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
- - - - -

Sequencing of Plasmids


  • Please log in to reply
2 replies to this topic

#1 Trinity

Trinity

    member

  • Active Members
  • Pip
  • 13 posts
0
Neutral

Posted 01 February 2005 - 02:44 AM

Hi! How do you sequence an insert in a Plasmid? Do you make a nick? How? Via Enzymes or in an ultrasoundbath?
How much ng of the plasmid do you use for the squencing reaction?

#2 badcell

badcell

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 94 posts
1
Neutral

Posted 01 February 2005 - 03:00 AM

You can use the supercoiled plasmid, you don't need to cut it. As few as 10-50 ng are enough, but I usually add 1 ul straight of the miniprep, without even OD'ed it . ;)
Science is a wonderful thing if one does not have to earn one's living at it
(A.Einstein)

#3 george@CASE

george@CASE

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 60 posts
0
Neutral

Posted 02 February 2005 - 10:51 AM

badcell is right. no need to do anything with the plasmid. Sequencing is like a PCR reaction only with just one primer.




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

©1999-2013 Protocol Online, All rights reserved.