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Team:University of Lethbridge/Notebook/GeneralLabJune
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====Nathan Puhl, Sebastian, Alix==== | ====Nathan Puhl, Sebastian, Alix==== | ||
Subcultured I13522 into liquid LB + amp for plasmid mini prep | Subcultured I13522 into liquid LB + amp for plasmid mini prep | ||
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| + | ===June 26, 2008=== | ||
| + | ====Nathan Puhl, Sebastian, Alix==== | ||
| + | Transformed RFP complete (BBa_J5526), pLACI-|TetR (BBa_I730002), Double T (BBa_B0015) | ||
| + | |||
| + | Plasmid mini-prepped GFP complete (BBa_I13522). | ||
Revision as of 02:40, 27 June 2008
Contents |
June 6 2008
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.
June 10 2008
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!)
June 11 2008
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.
June 16 2008
Nathan Puhl, Munima, Christa, Sebastian, Roxanne
Plated cheZ knockout strain from glycerol on LB + amp and Plain LB to assess viability and antibiotic resistance at 37 C overnight.
Transformed supercompetent cells with basic biobrick vector pSB1A7 (ampicillin resistance)
-50 uL of 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
June 17 2008
Munima, Christa, Nathan Puhl
Checked plates
-cheZ knockout strain viable on LB, no growth on LB + amp - Good -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
June 18 2008
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.
June 19 2008
Nathan Puhl, Alix, Munima, Christa, Roxanne
Ran plasmids on 1% agarose gel with High range ladder
June 24 2008
Nathan Puhl, Alix
Streaked BBa_I13522 (TetR repressed GFP) onto LB + amp from last year's glycerol stock.
June 25, 2008
Nathan Puhl, Sebastian, Alix
Subcultured I13522 into liquid LB + amp for plasmid mini prep
June 26, 2008
Nathan Puhl, Sebastian, Alix
Transformed RFP complete (BBa_J5526), pLACI-|TetR (BBa_I730002), Double T (BBa_B0015)
Plasmid mini-prepped GFP complete (BBa_I13522).
