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From 2010.igem.org

	Sunday	Monday	Tuesday	Wednesday	Thursday	Friday	Saturday
Week 1	-	6/28/2010	6/29/2010	6/30/2010	7/1/2010	7/2/2010	-
Week 2	-	-	-	7/7/2010	7/8/2010	7/9/2010	7/10/2010
Week 3	-	7/12/2010	-	-	-	-	-

Oil Sands Lab Notebook

This team includes Ann Lesnefsky, Bryce Rajabian, and Kilho Lee.

6/28/2010

Pseudomonas putida KT2440 Antibiotic Resistance

• Protocol for starting overnight culture from -80°C freezer
• All protocols can also be found in the Protocols section of the Notebook

6/29/2010

Pseudomonas putida KT2440 Antibiotic Resistance

• Protocol for making frozen stocks

6/29/2010 and 6/30/2010

Pseudomonas putida KT2440 Antibiotic Resistance

• Protocol for P. putida KT2440 antibotic resistance

7/1/2010

Literature Review Napthenic Acids Composition & HPLC Analysis

• Literature Review of the composition of napthenic acids in oil sands

7/2/2010

• Literature Review of HPLC columns for analysis of napthenic acids

Pseudomonas putida KT2440 Antibiotic Resistance

• Results

7/7/2010

Ann

Biobrick Transformation with Alex and Jennifer

• Made CaCl2 solution, and 100 mg/mL amphicilin stock solution according to the media section of the wiki
• Started an overnight culture of DH5alpha according to the heat shock transformation protocol
  • A 12 mL culture was started because we were multiplying the protocol by 4

7/8/2010

Ann

Biobrick Transformation with Marc, Katie and Audra according to the heat shock transformation protocol

• Started culture for biobrick transformation from over night at 2:30
• At 5:00pm the cultures had overgrown to an OD600 of 1.2
  • A a 1:3 dilution of cells was performed and the cells were allowed to go through another doubling period of 20 minutes
• The OD600 was measured again and found to be around 0.500
• the cultures were placed on ice at 5:30 and allowed to chill for 20 minutes
• The washings were performed and the comp cells were resuspended in the residual CaCl2 only
• The OD600 of the comp cells without the glycerol was above 0.3 therefore the comp cells were concentrated enough
• The heat shock was performed for the following biobricks
  • BBa_179015
  • BBa_179005
  • BBa_145001
  • BBa_117008
  • BBa_117002
  • BBa_103006
• The cultures were placed in the 30C shaker to grow up for an hour at 30C at 9:00pm
• The cultures were plated at 10:00pm

7/9/2010

Ann

Biobrick Transformation with Josh, Prae, Charlie according to the miniprep protocol

• After 9 hours of growing at 37C, the plates did not have any colonies on them
• After 22 hours of growth plates that had grown had clear colonies
  • The positive and negative controls grew out accordingly
  • Only BBa_179015 and BBa_179005 had colonies (both from plate 1 and none from plate 2...)
    • BBa_179015 had over 100 colonies
    • BBa_179005 had 2 colonies
• 5 mL overnight cultures were started with 100 mg/mL amp at 8pm from a single colony on each plate

7/10/2010

Ann

Biobrick Transformation with Marcus, Bryce, Kilho, Josh, Charlie, Jeremy

Miniprep

• Made frozen stocks from overnight culture of miniprep
• Performed miniprep on BBa_179015 and BBa_179005
• Measured DNA concentration with the Nanodrop
  • BBa_179015-11 ng/mL
  • BBa_179005-40 ng/mL

Digest

• Ran according to the digest protocol in the protocol section
  • Cut parts with EcoRI and PstI
• Digested at 37C for 30 minutes

Gel

• Ran according to the protocol in the protocol section
• Used 8 well comb instead of 13 well comb so the samples were too dilute too see

7/12/2010

Ann

Biobrick Transformation with Jeremy

Gel

• Reran gel from 7/10/2010

Lane 1-Invitrogen 1 kb plus ladder

Lane 2-Digested Bba_179015

• Ran out of undigested Bba_179015

Lane 3-Digested Bba_179005

Lane 4-Uncut miniprep plasmid Bba_179005