Team:University of Lethbridge/Notebook/GeneralLabSeptember

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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 pLac I culture. -Ran plasmids on a 1% agarose gel. -Oops, sub-culture it again.