Team:University of Lethbridge/Notebook/GeneralLabJune

From 2008.igem.org

(Difference between revisions)
(Nathan, ALix, Sebastian, Munima, Roxanne, Crista)
Line 98: Line 98:
Trasformed BBa_J24679( RBS + LacI), BBa_P0440 (RBS+TetR+T10+T12), leftover DNA from Double T (June 26, 2008), and 1 uL of pSB1A7 plasmid (June 18).
Trasformed BBa_J24679( RBS + LacI), BBa_P0440 (RBS+TetR+T10+T12), leftover DNA from Double T (June 26, 2008), and 1 uL of pSB1A7 plasmid (June 18).
-
Protocol changes   
+
Protocol changes:    
   -3 uL of DNA
   -3 uL of DNA
   -spin down 200 uL of cells and resuspend in 100 uL of LB; plate on LB + amp
   -spin down 200 uL of cells and resuspend in 100 uL of LB; plate on LB + amp

Revision as of 00:53, 3 July 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

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


June 17 2008

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


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

PSB1A7 plasmid.jpg

plasmid is ~15 ng/uL


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 BBa_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).

June 27, 2008

Nathan Puhl, Alix, Munima

No colonies on any plates. Will try again next week.

July 1, 2008

Nathan Puhl, Andrew

Made 500 mL of LB agar + amp and 500 mL of Liquid LB

July 2, 2008

Nathan, ALix, Sebastian, Munima, Roxanne, Crista

Trasformed BBa_J24679( RBS + LacI), BBa_P0440 (RBS+TetR+T10+T12), leftover DNA from Double T (June 26, 2008), and 1 uL of pSB1A7 plasmid (June 18).

Protocol changes:

  -3 uL of DNA
  -spin down 200 uL of cells and resuspend in 100 uL of LB; plate on LB + amp