Difference between revisions of "User:Daniel Goodman/Notebook/Cluzel/2010/04/05"

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(PZE-12-CFP vector from JM Kim)
(PZE-12-CFP vector from JM Kim)
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{| class="wikitable" border="1"
 
{| class="wikitable" border="1"
 
|-
 
|-
 +
! Symbol
 
!  Media
 
!  Media
 
!  Plasmid
 
!  Plasmid
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!  Why
 
!  Why
 
|-
 
|-
 +
|  A
 
|  LA/Amp
 
|  LA/Amp
 
|  PZE-12-CFP
 
|  PZE-12-CFP
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|  To get comp cells w/ plasmid
 
|  To get comp cells w/ plasmid
 
|-
 
|-
 +
|  B
 
|  LA
 
|  LA
 
|  PZE-12-CFP
 
|  PZE-12-CFP
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|  To check that my competent cells are alive
 
|  To check that my competent cells are alive
 
|-
 
|-
 +
|  C
 
|  Spec/AMP
 
|  Spec/AMP
 
|  PZS4-ATTP
 
|  PZS4-ATTP
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|  To check that my competent cells transform
 
|  To check that my competent cells transform
 
|-
 
|-
 +
|  D
 
|  LA/AMP
 
|  LA/AMP
 
|  PZE-12-CFP
 
|  PZE-12-CFP
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Protocol steps on page 1.119 - 1.122 of Molecular Cloning, Volume 1 book
 
Protocol steps on page 1.119 - 1.122 of Molecular Cloning, Volume 1 book
*need GYT medium
+
* used straight glycerol instead of GYT
*need SOC medium (1 ml per reaction)
+
* used SOC medium (1 ml per reaction)
* 10 ng of DNA per experiment  
+
* 1 ul of DNA per experiment (approx 52 ng for my plasmid, did not nanodrop Jeff's spec plasmid)
 +
*  1.8 kV, 1 second shock
  
  

Revision as of 14:37, 5 April 2010

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Prepping Plasmid for Electroporation

PZE-12-CFP vector from JM Kim

ng/ul 260/280 260/230
56.3 ng 1.92 1.62

Doing 4 electroporations, in order to ensure competent cells made last week work correctly.

Symbol Media Plasmid Cells Why
A LA/Amp PZE-12-CFP My competent MG1655 To get comp cells w/ plasmid
B LA PZE-12-CFP My competent MG1655 To check that my competent cells are alive
C Spec/AMP PZS4-ATTP My competent MG1655 To check that my competent cells transform
D LA/AMP PZE-12-CFP Jeff's competent MG1655 To check that my plasmid works/is correct

Protocol steps on page 1.119 - 1.122 of Molecular Cloning, Volume 1 book

  • used straight glycerol instead of GYT
  • used SOC medium (1 ml per reaction)
  • 1 ul of DNA per experiment (approx 52 ng for my plasmid, did not nanodrop Jeff's spec plasmid)
  • 1.8 kV, 1 second shock