Team:University of Lethbridge/Notebook/GeneralLabJune

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Sebastian, John and Roxanne
Prepared 1L of semi-solid media following the procedure found on OpenWetWare. -10g peptone (substituted for tryptone) -10g Agar -10g NaCl -5g Yeast Extract

Stored media in fridge.

Christa, Munima, Roxanne, and Sebastian
Prepared 1L of liquid media following the procedure found on OpenWetWare. -10g peptone (substituted for tryptone) -10g NaCl -5g Yeast Extract

Poured 36 culture test tubes, and the remainder was left in a 1L Erlenmeyer flask. Stored both in fridge.

Christa
Made an inventory of iGEM 2007 parts in Wieden -80 freezer inventory.xls

Roxanne
Defrosted the -20 freezer in the teaching lab for iGEM use (with a little help from my friends!)

Sebastian, Munima, Roxanne, Christa
Poured 8 minimal media (labeled control - with blue sharpie) plates and 14 amp plates (labeled amp - with red sharpie) stored in the 4 C fridge. Amp concentration is always 50ug/mL.

Nathan Puhl, Munima, Christa, Sebastian, Roxanne
Transformed supercompetent cells with basic biobrick vector pSB1A7 (ampicillin resistance). -50 uL of DH5-alpha cells, 0.85 uL 2-mercaptoethanol, 1 uL of plasmid dissolved with 15 uL of ddH2O -30 min on ice -45 s at 42 C -2 min on ice -Add 0.9 mL of LB shaker incubate at 225 RPM and 37 C

Munima, Christa, Nathan Puhl
Checked plates -Only one colony from transformation, not very good efficiency, don't know why because too many possibilities, most likely amount of DNA due to inability to quantify plasmid from iGEM plates -Subcultured colony in liquid LB + amp -Plate 200 uL on LB + amp at 37 C overnight

Munima, Christa, Alix, Nathan Puhl
Made glycerol stock of pSB1A7 transformed E. coli

Extracted plasmid from transformed E. coli using the Eppendorf FastPlasmid minikit and stored 4 aliquots of 25 uL in the -20 C freezer.

Nathan Puhl, Alix, Munima, Christa, Roxanne
Ran plasmids on 1% agarose gel with High range ladder



plasmid is ~15 ng/uL

Nathan Puhl, Alix
Streaked BBa_I13522 (TetR repressed GFP) onto LB + amp from last year's glycerol stock.

Nathan Puhl, Sebastian, Alix
Subcultured BBa_I13522 into liquid LB + amp for plasmid mini prep

Nathan Puhl, Sebastian, Alix
Transformed BBa_J5526 (RFP complete), BBa_I730002 (pLACI-|TetR), and BBa_B0015 (Double T).

Plasmid mini-prepped GFP complete (BBa_I13522).

Nathan Puhl, Alix, Munima
No colonies on any plates. Will try again next week.