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
- - - - -

recognition sites and cloning?


  • Please log in to reply
2 replies to this topic

#1 boa

boa

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 01 October 2012 - 08:12 PM

Hello all, I have absolutely no clue how to start this problem and I would like to have some cues or just something to chew on...thank you!!
I know what a plasmid is , I know what sticky ends (cohesive extremities) are but I dont know how the multiple cloning sites in this plasmid are located so im a bit lost!

Question: U must clone a DNA fragment having Sau3A sticky ends.
The plasmid to be used for the cloning experiment has the following MCS
EcoR1, BamH1, Xba1, HindIII

*Describe how you would do the cloning experiment.

Restriction enzyme recognition sites
EcoR1 = GAATTC (the enzyme cuts after the G)
BamH1 = GGATTC (the enzyme cust after the first G)
Xba1 = TCTAGA (the enzyme cuts after the first T)
HindIII = AAGCTT (the enzyme cuts after the first A)
Sau3A = GATC (the enzyme cuts before the G)

#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,512 posts
371
Excellent

Posted 01 October 2012 - 11:39 PM

Your BamH1 is wrong - should be GGATCC - which will help you in your cloning (think which ends will be compatible with Sau3a).

As this is homework, I'm not giving a full answer, you need to provide a bit more input so that we can help you learn rather than just having us supply the answer.

#3 susnata

susnata

    member

  • Active Members
  • Pip
  • 10 posts
0
Neutral

Posted 26 July 2013 - 01:45 AM

i think you already got the answer... it's called "compatible cohesive ends"




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

©1999-2013 Protocol Online, All rights reserved.