User:Eric Ma/Notebook/MICB323 Lab Book/2009/01/27

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Part 1: Transformation using ligation reaction and one shot INVaF' CaCl2 prepared competent cells

 * Protocol followed just as outlined in the Lab Manual pg 33

Part 2: Transformation via electroporation using supplied p421 plasmid

 * Note: Sunny will do the p421 and CaCl2
 * Protocol followed as outlined in Lab Manual pages 37-38

Plates from Part 1

 * These are CaCl2 transformed cells using INVaF' competent cells and ligation reaction plasmid.
 * 3 plates:

Plates from Parts 2 and 3
These plates come from electroporation using the p421 plasmid. See this [[Media:090127_Plates_Summary.xlsx|Excel spreadsheet]] to see how calculations were done.

Summary
I have a few questions: Plates selected
 * 1) Plating went well and smoothly.
 * 2) Electroporation is surprisingly easy! Just push that button. "That was easy!"
 * 1) What is the purpose of the pre-incubation plating and post-incubation plating of the original culture? Is it some control of sorts?
 * 2) The purpose of plating on LB alone - to get a feel for the total number of cells present?
 * 3) We can then take total number of transformants (i.e. from the LB-AMP + X-gal) divided by total number of cells used - get a feel for efficiency of transformation?
 * 1) Selected pure white and pure blue colonies from ligation reaction transformants. Prospective ones are labeled on the "Part 1" plate with "W" and "B" respectively.
 * 2) Selected pure white colonies from p421 transformants. Pick them from the "Part 2(a)" plate on Monday.


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