Team:WashU/Notebook/MolecularBiology

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==Week of 6/28==
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===7/1===
 +
Plated The Megax BH10B strain and Strain 8 from the cohen lab onto an amp plate to check that the Megax BH10B strain is killed
 +
==Week of 7/12==
 +
===7/13===
 +
Minipreped all the cultures and nanodropped them
 +
 +
Ran 3 PCR reactions (@ 58,60, and 62 C) with primers p1 and p2 in order to attach the kozak onto the YFP
 +
 +
Enzyme digested Nat (bglII and EcoRI), Kan (BamHI and EcoRI), and Promoter (EcoRI).  Did 2 reactions each with ~500 ng DNA
 +
 +
==Week of 7/19==
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Gel from July 13th
 +
 +
Lanes from top to bottom:
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1)  Ladder
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2)  5uL of 58 degree YFP + 1uL of loading dye
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3)  5uL of 60 degree YFP + 1uL of loading dye
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4)  5uL of 62 degree YFP + 1uL of loading dye
 +
5)  5uL of KanMx4 1 + 1uL of loading dye
 +
6)  5uL of KanMx4 2+ 1uL of loading dye
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7)  5uL of NatMx4 1 + 1uL of loading dye
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8)  5uL of NatMx4 2+ 1uL of loading dye
 +
9)  5uL of Promoter 1 + 1uL of loading dye
 +
10) 5uL of Promoter 2 + 1uL of loading dye
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 +
[[Image:WashU7-24Gel.png]]
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===7/15===
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Gel reads from left to right as follows:
 +
Dna Ladder;
 +
NatMX4-Promoter construct 2x;
 +
KanMX4 plasmid backbone 2x.
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DNA ladder  information is here at http://www.neb.com/nebecomm/products/productN3232.asp
 +
 +
[[Image:WashU7-15Gel1.PNG]]
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[[Image:WashU7-15Gel2.PNG]]
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 +
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==Week of 7/26==
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==Week of 8/2==
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==Week of 8/9==
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==Week of 8/16==
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 +
==Week of 8/23==
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==Week of 8/30==

Revision as of 06:15, 8 October 2010


Week of 6/28

7/1

Plated The Megax BH10B strain and Strain 8 from the cohen lab onto an amp plate to check that the Megax BH10B strain is killed

Week of 7/12

7/13

Minipreped all the cultures and nanodropped them

Ran 3 PCR reactions (@ 58,60, and 62 C) with primers p1 and p2 in order to attach the kozak onto the YFP

Enzyme digested Nat (bglII and EcoRI), Kan (BamHI and EcoRI), and Promoter (EcoRI). Did 2 reactions each with ~500 ng DNA

Week of 7/19

Gel from July 13th

Lanes from top to bottom: 

1)   Ladder
2)   5uL of 58 degree YFP + 1uL of loading dye
3)   5uL of 60 degree YFP + 1uL of loading dye
4)   5uL of 62 degree YFP + 1uL of loading dye
5)   5uL of KanMx4 1 + 1uL of loading dye
6)   5uL of KanMx4 2+ 1uL of loading dye
7)   5uL of NatMx4 1 + 1uL of loading dye
8)   5uL of NatMx4 2+ 1uL of loading dye
9)   5uL of Promoter 1 + 1uL of loading dye
10) 5uL of Promoter 2 + 1uL of loading dye

WashU7-24Gel.png

7/15

Gel reads from left to right as follows: 

Dna Ladder; 
NatMX4-Promoter construct 2x; 
KanMX4 plasmid backbone 2x.

DNA ladder information is here at http://www.neb.com/nebecomm/products/productN3232.asp

File:WashU7-15Gel1.PNG

File:WashU7-15Gel2.PNG


Week of 7/26

Week of 8/2

Week of 8/9

Week of 8/16

Week of 8/23

Week of 8/30

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