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)
Line 19: Line 19:
{| 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
Line 46: Line 51:
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 17:37, 5 April 2010


Cluzel Lab Notebook
Daniel Goodman
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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



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