Lec9_W11

Lec9_W11 - Transformation efficiency This a calculation...

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Transformation efficiency This a calculation that is done to determine how competent cells are You need to know exactly how much plasmid you added to the competent cells in order to do the calculations
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You took 1 ng (in 5 μ l) of pGEM and added it to 45 μ l of competent cells You then added 950 μ l of LB to the cells,so you had 1 ng of pGEM + an unkown number of cells in 1 ml of volume You plated 100 μ l or 1/10 th of that total on a plate. Let’s say you got 2000 colonies on that plate. That means that each of those colonies arose from one cell that was transformed by pGEM. By extrapolation, we could estimate that if you had plated the whole ml of LB, you would have gotten 20,000 colonies. That means that you had 20,000 cells that were transformed by the addition of 1 ng pGEM. If we had increased everything by 1000-fold, you would have gotten 20,000,000 cells transformed by 1 μ g of pGEM. Your transformation efficiency is said to be 2 x10 7 colonies / μ g of pGEM
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Vibrio fischeri DNA isolation Create genomic library in pGEM and DH5 α Screen for expression of bioluminescence genes Pick glowing colony,
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This note was uploaded on 04/10/2011 for the course BIMM 101 taught by Professor Butler during the Spring '08 term at UCSD.

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Lec9_W11 - Transformation efficiency This a calculation...

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