Team:University of Lethbridge/Notebook/GeneralLabJune

From 2008.igem.org

(Difference between revisions)
m
 
(16 intermediate revisions not shown)
Line 1: Line 1:
 +
[[Team:University_of_Lethbridge/Notebook|Back to The University of Lethbridge Main Notebook]]
 +
===June 6 2008===
===June 6 2008===
-
====Sebastian and Roxanne ====
+
====Sebastian, John 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 (substituted for tryptone)
  -10g peptone (substituted for tryptone)
Line 8: Line 10:
Stored media in fridge.
Stored media in fridge.
 +
===June 10 2008===
===June 10 2008===
====Christa, Munima, Roxanne, and Sebastian====
====Christa, Munima, Roxanne, and Sebastian====
Prepared 1L of liquid media following the procedure found on OpenWetWare.
Prepared 1L of liquid media following the procedure found on OpenWetWare.
-
  -10g peptone
+
  -10g peptone (substituted for tryptone)
  -10g NaCl
  -10g NaCl
  -5g Yeast Extract
  -5g Yeast Extract
Poured 36 culture test tubes, and the remainder was left in a 1L Erlenmeyer flask. Stored both in fridge.
Poured 36 culture test tubes, and the remainder was left in a 1L Erlenmeyer flask. Stored both in fridge.
-
 
====Christa====
====Christa====
Line 25: Line 27:
====Roxanne====
====Roxanne====
Defrosted the -20 freezer in the teaching lab for iGEM use (with a little help from my friends!)
Defrosted the -20 freezer in the teaching lab for iGEM use (with a little help from my friends!)
 +
===June 11 2008===
===June 11 2008===
====Sebastian, Munima, Roxanne, Christa====
====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.
+
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===
===June 16 2008===
====Nathan Puhl, Munima, Christa, Sebastian, Roxanne====
====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 [http://partsregistry.org/wiki/index.php?title=Part:pSB1A7 pSB1A7] (ampicillin resistance).
-
Transformed supercompotent 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
-
   -50 uL of cells, 0.85 uL 2-mercaptoethanol, 1 uL of plasmid dissolved with 15 uL of ddH2O
+
   -30 min on ice
   -30 min on ice
   -45 s at 42 C
   -45 s at 42 C
   -2 min on ice
   -2 min on ice
   -Add 0.9 mL of LB shaker incubate at 225 RPM and 37 C
   -Add 0.9 mL of LB shaker incubate at 225 RPM and 37 C
 +
===June 17 2008===
===June 17 2008===
====Munima, Christa, Nathan Puhl====
====Munima, Christa, Nathan Puhl====
Checked plates
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      
-
   -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
+
    likely amount of DNA due to inability to quantify plasmid from iGEM plates
   -Subcultured colony in liquid LB + amp
   -Subcultured colony in liquid LB + amp
   -Plate 200 uL on LB + amp at 37 C overnight
   -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
 +
 +
[[Image:pSB1A7 plasmid.jpg|500 px]]
 +
 +
plasmid is ~15 ng/uL
 +
 +
 +
===June 24 2008===
 +
====Nathan Puhl, Alix====
 +
Streaked [http://partsregistry.org/wiki/index.php?title=Part:BBa_I13522 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 [http://partsregistry.org/wiki/index.php?title=Part:BBa_J5526 BBa_J5526] (RFP complete), [http://partsregistry.org/wiki/index.php?title=Part:BBa_I730002 BBa_I730002] (pLACI-|TetR), and [http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015 BBa_B0015] (Double T).
 +
 +
Plasmid mini-prepped GFP complete (BBa_I13522).
 +
 +
 +
===June 27, 2008===
 +
====Nathan Puhl, Alix, Munima====
 +
No colonies on any plates.  Will try again next week.

Latest revision as of 16:31, 25 August 2008

Back to The University of Lethbridge Main Notebook

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 [http://partsregistry.org/wiki/index.php?title=Part:pSB1A7 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 [http://partsregistry.org/wiki/index.php?title=Part:BBa_I13522 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 [http://partsregistry.org/wiki/index.php?title=Part:BBa_J5526 BBa_J5526] (RFP complete), [http://partsregistry.org/wiki/index.php?title=Part:BBa_I730002 BBa_I730002] (pLACI-|TetR), and [http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015 BBa_B0015] (Double T).

Plasmid mini-prepped GFP complete (BBa_I13522).


June 27, 2008

Nathan Puhl, Alix, Munima

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

Retrieved from "http://2008.igem.org/Team:University_of_Lethbridge/Notebook/GeneralLabJune"