Team:University of Lethbridge/Notebook/GeneralLabJune

From 2008.igem.org

(Difference between revisions)
(→June 10 2008)
Line 2: Line 2:
====Sebastian and Roxanne ====
====Sebastian and Roxanne ====
Prepared 1L of semi-solid media following the procedure found on OpenWetWare.
Prepared 1L of semi-solid media following the procedure found on OpenWetWare.
-
  -10g peptone
+
  -10g peptone (substituted for tryptone)
  -10g Agar
  -10g Agar
  -10g NaCl
  -10g NaCl
  -5g Yeast Extract
  -5g Yeast Extract
-
Semi-solid Media was autoclaved and stored in the iGEM fridge for making plates at a later date.
+
Stored media in fridge.
===June 10 2008===
===June 10 2008===
Line 16: Line 16:
  -5g Yeast Extract
  -5g Yeast Extract
-
36 culture test tubes were poured, and the remainder was left in a 1L Erlenmeyer flask.
+
Poured 36 culture test tubes, and the remainder was left in a 1L Erlenmeyer flask. Stored both in fridge.
-
Both were autoclaved and stored in the iGEM fridge.
+
 
====Christa====
====Christa====
-
Took and inventory of iGEM 2007 parts in Wieden -80 freezer
+
Made an inventory of iGEM 2007 parts in Wieden -80 freezer
 +
[[inventory.xls]]
====Roxanne====
====Roxanne====
Line 28: Line 29:
====Sebastian, Munima, Roxanne, Christa====
====Sebastian, Munima, Roxanne, Christa====
-
Poured 8 minimal media (labeled control- with blue sharpie) plates and 14 ampicillin plates (labeled ampicillin- with red sharpie)with aseptic technique and stored in the 4C fridge. Ampicillin concentration is 50ug/mL according to the Topp and Gallivan paper.
+
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 supercompotent 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 possiblilities, most likely amount of DNA due to inability to quantifiy plasmid from iGEM plates
 +
  -Subcultured colony in liquid LB + amp
 +
  -Plate 200 uL on LB + amp at 37 C overnight

Revision as of 23:35, 17 June 2008

Contents

June 6 2008

Sebastian 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
-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 supercompotent 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 possiblilities, most likely amount of DNA due to inability to quantifiy plasmid from iGEM plates
 -Subcultured colony in liquid LB + amp
 -Plate 200 uL on LB + amp at 37 C overnight
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