Team:University of Lethbridge/Notebook/GeneralLabJune

From 2008.igem.org

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[[Team:University_of_Lethbridge/Notebook|Back to The University of Lethbridge Main Notebook]]
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===June 6 2008===
===June 6 2008===
====Sebastian, John and Roxanne ====
====Sebastian, John and Roxanne ====
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===June 16 2008===
===June 16 2008===
====Nathan Puhl, Munima, Christa, Sebastian, Roxanne====
====Nathan Puhl, Munima, Christa, Sebastian, Roxanne====
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Plated cheZ knockout strain from glycerol on LB + amp and Plain LB to assess viability and antibiotic resistance at 37 C overnight.
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Transformed supercompetent cells with basic biobrick vector [http://partsregistry.org/wiki/index.php?title=Part:pSB1A7 pSB1A7] (ampicillin resistance).
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   -50 uL of DH5-alpha cells, 0.85 uL 2-mercaptoethanol, 1 uL of plasmid dissolved with 15 uL of ddH2O
-
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
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   -30 min on ice
   -30 min on ice
   -45 s at 42 C
   -45 s at 42 C
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====Munima, Christa, Nathan Puhl====
====Munima, Christa, Nathan Puhl====
Checked plates
Checked plates
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  -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 possibilities, most       
     likely amount of DNA due to inability to quantify plasmid from iGEM plates
     likely amount of DNA due to inability to quantify plasmid from iGEM plates
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[[Image:pSB1A7 plasmid.jpg|500 px]]
[[Image:pSB1A7 plasmid.jpg|500 px]]
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plasmid is ~15 ng/uL
===June 24 2008===
===June 24 2008===
====Nathan Puhl, Alix====
====Nathan Puhl, Alix====
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Streaked BBa_I13522 (TetR repressed GFP) onto LB + amp from last year's glycerol stock.
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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===
===June 25, 2008===
====Nathan Puhl, Sebastian, Alix====
====Nathan Puhl, Sebastian, Alix====
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Subcultured I13522 into liquid LB + amp for plasmid mini prep
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Subcultured BBa_I13522 into liquid LB + amp for plasmid mini prep
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===June 26, 2008===
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====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).
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Plasmid mini-prepped GFP complete (BBa_I13522).
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 +
 
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===June 27, 2008===
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====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.