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
Edited by Agape, 24 July 2011 - 05:49 AM.
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Glycerol stock
Started by Agape, Jul 24 2011 05:48 AM
3 replies to this topic
#1
Agape
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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
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
#2
Adrian K
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Posted 24 July 2011 - 07:36 AM
Agape, on 24 July 2011 - 05:48 AM, said:
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
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
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
..."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
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Posted 24 July 2011 - 08:21 AM
Adrian K, on 24 July 2011 - 07:36 AM, said:
Agape, on 24 July 2011 - 05:48 AM, said:
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
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
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
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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.
