User:Eric Ma/Notebook/MICB323 Lab Book/2009/02/03: Difference between revisions

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==Part 1: Plasmid Extraction==
* Electroporation transformed: No growth.
* CaCl<sub>2</sub> untransformed: Growth.
* CaCl<sub>2</sub> transformed: Growth.
* I will use Sunny's culture's DNA.
<br>
*Entire ON culture - spin @8000rpm @3min. Performed split up into two spins in same tubes.


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Revision as of 14:21, 3 February 2009

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Pre-Lab Calculations

  • Done as listed in this spreadsheet.
  • Volumes to be used:
Variable Tube # pDNA (µL) EcoR1 (µL) 10X React 3 (µL) 500 mM NaCl (µL) 500 mM Tris-HCl (µL) 100 mM MgCl2 (µL) dH2O (µL) Total Volume (µL)
[enzyme] 1 0.2 0.5 2 0 0 0 17.3 20
[enzyme] 2 0.2 1 2 0 0 0 16.8 20
[enzyme] 3 0.2 1.5 2 0 0 0 16.3 20
[enzyme] 4 0.2 2 2 0 0 0 15.8 20
[NaCl] 1 0.2 0.5 2 1 0 0 16.3 20
[NaCl] 2 0.2 0.5 2 2 0 0 15.3 20
[NaCl] 3 0.2 0.5 2 3 0 0 14.3 20
[NaCl] 4 0.2 0.5 2 4 0 0 13.3 20
  • Calculation table:
Component Tube [NaCl] in 10X React 3 (mM) [NaCl] Provided (mM) [NaCl] Wanted (10X) (mM) Extra [NaCl] needed (mM) 500 mM NaCl (µL)
NaCl 1 500 500 1000 500 1
NaCl 2 500 500 1500 1000 2
NaCl 3 500 500 2000 1500 3
NaCl 4 500 500 2500 2000 4

Part 1: Plasmid Extraction

  • Electroporation transformed: No growth.
  • CaCl2 untransformed: Growth.
  • CaCl2 transformed: Growth.
  • I will use Sunny's culture's DNA.


  • Entire ON culture - spin @8000rpm @3min. Performed split up into two spins in same tubes.


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