Team:University of Lethbridge/Notebook/GeneralLabSeptember

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[[Team:University_of_Lethbridge/Notebook|Back to The University of Lethbridge Main Notebook]]
[[Team:University_of_Lethbridge/Notebook|Back to The University of Lethbridge Main Notebook]]
 +
 +
===August 31, 2008===
 +
====Nathan Puhl, Roxanne====
 +
-Transformed DH5a cells with the biobrick part R0011 (pLacI) obtained from the 2007 iGEM parts. Plated on LB + amp agar plates.
 +
 +
 +
===September 1, 2008===
 +
====Roxanne====
 +
-Picked 3 representative colonies and incubated them in LB+amp media overnight at 37.0C.
 +
 +
 +
===September 2, 2008===
 +
====Roxanne====
 +
-plasmid prepped the pLacI cells which had been incubated the night before. Ran plasmids on a 1% Agarose Gel at 100V for 27 minutes
 +
 +
====Nathan Puhl====
 +
-Visualized the Gel and Glycerol Stocked the Cells.
 +
 +
 +
===September 3, 2008===
 +
====Roxanne, Nathan Puhl====
 +
-Performed a Restriction Digest of I13504 (RBS+GFP+dT) and R0011 (pLacI recombinant)
 +
 +
-10 uL template DNA
 +
-0.5 uL Restriction Enzyme #1
 +
-0.5 uL Restriction Enzyme #2
 +
-2 uL NEB 2
 +
-7 uL ddH2O
 +
_________
 +
20 uL Reaction overnight at 37.0C
 +
 +
===Mid-September, 2008===
 +
====Roxanne, Nathan Puhl====
 +
-Have been working on Cloning the Reporter System. Transformation worked, however, the lack of Red colonies on the RFP plates and sub-culture have led us to believe that something didn't work somewhere along the way.
 +
 +
 +
===September 13, 2008===
 +
====Roxanne====
 +
-Streaked Plates of RFP sub, TetR sub and pLacI
 +
 +
 +
===September 14, 2008===
 +
====Roxanne====
 +
-Picked colonies from the RFP sub, TetR sub and pLacI plates. Incubating overnight at 37.0C in LB media + amp.
 +
 +
 +
===September 17, 2008===
 +
====Nathan Puhl====
 +
-Plasmid prepped the RFP sub, TetR sub and pLacI cultures.
 +
 +
====Nathan Puhl, Roxanne====
 +
-Restriction digest of the RFP sub and TetR sub with XbaI and PstI
 +
-Restriction Digest of pLacI with SpeI and PstI.
 +
 +
 +
===September 18, 2008===
 +
====Nathan Puhl====
 +
-Ran a 1% agarose gel of the digestion products. RFP and TetR look good, however, pLacI is running high.
 +
 +
 +
===September 19, 2008===
 +
====Nathan Puhl, Roxanne====
 +
-repicked colonies from pLacI plates for subculture in LB media.
 +
 +
 +
===September 20, 2008===
 +
====Nathan Puhl, Roxanne====
 +
-plasmid prepped the pLacI culture.
 +
-Ran plasmids on a 1% agarose gel.
 +
-Oops, sub-culture it again.
 +
 +
 +
===September 21, 2008===
 +
====Roxanne====
 +
-plasmid prepped the pLacI culture.
 +
-Ran the gel of the plasmid prep products
 +
 +
[[Image:PLacIprep.jpg|350 px]]
 +
 +
 +
===September 22, 2008===
 +
====Roxanne====
 +
-Gel Extraction of pLacI x2, RFP sub, and TetR sub
 +
 +
[[Image:extraction.jpg|350 px]]
 +
 +
-Ran a gel of the extraction products, however, no product appears.
 +
 +
 +
===September 23, 2008===
 +
====Roxanne====
 +
-Ran the gel of the extraction products again.
 +
-1 uL of DNA on a 1% agarose gel @ 100V for 25 minutes, with a 1kb ladder.
 +
-The Kit ate my DNA again!!
 +
-Repicked colonies from the plate and incubated in LB+amp media overnight @ 37.0C
 +
 +
 +
===September 24, 2008===
 +
====Roxanne====
 +
-Took the tubes out of the shaker and put in fridge to be plasmid prepped at a later date.
 +
 +
 +
===September 27, 2008===
 +
====Roxanne====
 +
-plasmid prepped the pLacI, RFP and TetR genes
 +
-Ran the plasmids on a gel. RFP and TetR look good, pLacI is a little wonky, not comforteble with it
 +
-Setup a Restriction Digest of RFP and TetR, using XbaI and PstI.
 +
-Repicked te pLacI colony and incubated in LB+amp overnight @ 37.0C
 +
 +
 +
===September 28, 2008===
 +
====Roxanne====
 +
-Plasmid prepped the pLacI gene and put te plasmid in the freezer
 +
-Inactivated the enzymes from the RFP and TetR double digest
 +
 +
 +
===September 30, 2008===
 +
====Roxanne====
 +
-Testing the Enzymes we received from Fermentas which were sent without ice on the pLacI gene
 +
-Cut Half of my plasmid prep with the iGEM SpeI and PstI, and the other half with H-J's PstI and the iGEM SpeI.

Latest revision as of 02:30, 30 October 2008

Back to The University of Lethbridge Main Notebook

Contents

August 31, 2008

Nathan Puhl, Roxanne

-Transformed DH5a cells with the biobrick part R0011 (pLacI) obtained from the 2007 iGEM parts. Plated on LB + amp agar plates.


September 1, 2008

Roxanne

-Picked 3 representative colonies and incubated them in LB+amp media overnight at 37.0C.


September 2, 2008

Roxanne

-plasmid prepped the pLacI cells which had been incubated the night before. Ran plasmids on a 1% Agarose Gel at 100V for 27 minutes

Nathan Puhl

-Visualized the Gel and Glycerol Stocked the Cells.


September 3, 2008

Roxanne, Nathan Puhl

-Performed a Restriction Digest of I13504 (RBS+GFP+dT) and R0011 (pLacI recombinant) 

-10 uL template DNA
-0.5 uL Restriction Enzyme #1
-0.5 uL Restriction Enzyme #2
-2 uL NEB 2
-7 uL ddH2O
_________
20 uL Reaction overnight at 37.0C

Mid-September, 2008

Roxanne, Nathan Puhl

-Have been working on Cloning the Reporter System. Transformation worked, however, the lack of Red colonies on the RFP plates and sub-culture have led us to believe that something didn't work somewhere along the way.


September 13, 2008

Roxanne

-Streaked Plates of RFP sub, TetR sub and pLacI


September 14, 2008

Roxanne

-Picked colonies from the RFP sub, TetR sub and pLacI plates. Incubating overnight at 37.0C in LB media + amp.


September 17, 2008

Nathan Puhl

-Plasmid prepped the RFP sub, TetR sub and pLacI cultures.

Nathan Puhl, Roxanne

-Restriction digest of the RFP sub and TetR sub with XbaI and PstI -Restriction Digest of pLacI with SpeI and PstI.


September 18, 2008

Nathan Puhl

-Ran a 1% agarose gel of the digestion products. RFP and TetR look good, however, pLacI is running high.


September 19, 2008

Nathan Puhl, Roxanne

-repicked colonies from pLacI plates for subculture in LB media.


September 20, 2008

Nathan Puhl, Roxanne

-plasmid prepped the pLacI culture. -Ran plasmids on a 1% agarose gel. -Oops, sub-culture it again.


September 21, 2008

Roxanne

-plasmid prepped the pLacI culture. -Ran the gel of the plasmid prep products

PLacIprep.jpg


September 22, 2008

Roxanne

-Gel Extraction of pLacI x2, RFP sub, and TetR sub

Extraction.jpg

-Ran a gel of the extraction products, however, no product appears.


September 23, 2008

Roxanne

-Ran the gel of the extraction products again. -1 uL of DNA on a 1% agarose gel @ 100V for 25 minutes, with a 1kb ladder. -The Kit ate my DNA again!! -Repicked colonies from the plate and incubated in LB+amp media overnight @ 37.0C


September 24, 2008

Roxanne

-Took the tubes out of the shaker and put in fridge to be plasmid prepped at a later date.


September 27, 2008

Roxanne

-plasmid prepped the pLacI, RFP and TetR genes -Ran the plasmids on a gel. RFP and TetR look good, pLacI is a little wonky, not comforteble with it -Setup a Restriction Digest of RFP and TetR, using XbaI and PstI. -Repicked te pLacI colony and incubated in LB+amp overnight @ 37.0C


September 28, 2008

Roxanne

-Plasmid prepped the pLacI gene and put te plasmid in the freezer -Inactivated the enzymes from the RFP and TetR double digest


September 30, 2008

Roxanne

-Testing the Enzymes we received from Fermentas which were sent without ice on the pLacI gene -Cut Half of my plasmid prep with the iGEM SpeI and PstI, and the other half with H-J's PstI and the iGEM SpeI.

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