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

Glycerol stock


  • Please log in to reply
3 replies to this topic

#1 Agape

Agape

    member

  • Members
  • Pip
  • 4 posts
0
Neutral

Posted 24 July 2011 - 05:48 AM

I have worked on plasmid DNA isolation and cell retransformation and i was ask to prepare glycerol stock for recombinant strain and how to ensure it is functional of gene expression(refering to glycerol stock).

For preparation of glycerol stock: I chose -70oC with 15% glycerol.

However, i am not sure how to ensure that it is functional.

Any help will be appreciated :D

Edited by Agape, 24 July 2011 - 05:49 AM.


#2 Adrian K

Adrian K

    Legendary Graduate Beggar

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 708 posts
28
Excellent

Posted 24 July 2011 - 07:36 AM

I have worked on plasmid DNA isolation and cell retransformation and i was ask to prepare glycerol stock for recombinant strain and how to ensure it is functional of gene expression(refering to glycerol stock).

For preparation of glycerol stock: I chose -70oC with 15% glycerol.

However, i am not sure how to ensure that it is functional.

Any help will be appreciated :D


If your cell is E.coli, is ok.
Expecting the world to treat you fairly because you are a good person is like expecting the lion not to attack you because you are a vegetarian.

..."best of our knowledge, as far as we know this had never been reported before, though I can't possible read all the published journals on earth, but by perform thorough search in google, the keywords did not match any documents"...

"what doesn't kill you, makes you stronger"---Goddess Casandra reminds me to be strong

"It's all just DNA. Do it."---phage434

#3 Agape

Agape

    member

  • Members
  • Pip
  • 4 posts
0
Neutral

Posted 24 July 2011 - 08:21 AM


I have worked on plasmid DNA isolation and cell retransformation and i was ask to prepare glycerol stock for recombinant strain and how to ensure it is functional of gene expression(refering to glycerol stock).

For preparation of glycerol stock: I chose -70oC with 15% glycerol.

However, i am not sure how to ensure that it is functional.

Any help will be appreciated :D


If your cell is E.coli, is ok.


Thanks bro:) However, my prof wants me prove that it is functional. What can i do? btw, the recombinant cell should produce GFP by induction of IPTG. Any advices? (like should i prove the plasmid stability or whatsoever?)

#4 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,726 posts
399
Excellent

Posted 24 July 2011 - 04:52 PM

For functionality, the easiest way is to prepare plasmid from bacteria taken from your glycerol stock, and then express the protein encoded, detect by western blot.




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

©1999-2013 Protocol Online, All rights reserved.