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Team:Waterloo/Notebook
From 2011.igem.org
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*Frozen stock of GFP2 and lox made | *Frozen stock of GFP2 and lox made | ||
*Miniprep of GFP1 x4 replicates | *Miniprep of GFP1 x4 replicates | ||
| - | Gel extraction | + | *Gel extraction of IN1 and IN2 resulted in improved concentrations |
| + | *Transformation | ||
Revision as of 20:02, 27 July 2011
This is a template page. READ THESE INSTRUCTIONS.
You are provided with this team page template with which to start the iGEM season. You may choose to personalize it to fit your team but keep the same "look." Or you may choose to take your team wiki to a different level and design your own wiki. You can find some examples HERE.
You MUST have a team description page, a project abstract, a complete project description, a lab notebook, and a safety page. PLEASE keep all of your pages within your teams namespace.
| You can write a background of your team here. Give us a background of your team, the members, etc. Or tell us more about something of your choosing. | File:Waterloo logo.png 200px |
|
Tell us more about your project. Give us background. Use this is the abstract of your project. Be descriptive but concise (1-2 paragraphs) | File:Waterloo team.png Your team picture |
| Team Example |
| Home | Team | Official Team Profile | Project | Parts Submitted to the Registry | Modeling | Notebook | Safety | Attributions |
|---|
Notebook
You should make use of the calendar feature on the wiki and start a lab notebook. This may be looked at by the judges to see how your work progressed throughout the summer. It is a very useful organizational tool as well.
Wednesday July 6, 2011
- Received 3 of 4 sequences the previous week (IN1, IN2 and GFP2).
- A quick spin down UWAT014-3/UWAT014-2 using centrifuge.
- Resuspended DNA in 40ul of MQ water (Concentration: 2ug/40ul=1ug/20ul)
- Transformed into DH5-alpha (sequences in PUC57).
- grown overnight on ampicillin plates.
- Resuspension of PSB1C3 in the Spring 2011 distribution kit (Plate 1 well 3A). Contains BBa_J04450.
- Resuspension of PSB1C3 in 10ul of MQ water (aspirated), wait approximately 5 minutes.
- 1ul of resuspension was transformed into DH5-alpha. Grown overnight on plate.
Thursday July 7, 2011
- IN1 (amp), IN2 (amp), GFP2 (amp) and PSB1C3 (cm) broth cultures innoculated (3 each).
Friday July 8, 2011
- Frozen stock of IN1, IN2 and GFP2 in PUC57 and PSB1C3 backbone made.
- Miniprep for IN1, IN2, GFP2 and PUC57 completed:
| Sequences | IN1 | IN2 | GFP2 | PSB1C3 |
| 260/280 | 1.85 | 1.80 | 1.88 | 1.86 |
| ng/ul | 229.8 | 236.1 | 198.6 | 166.2 |
- GFP1 Sequence(588nt)in PUC57 received from Bio Basic Canada INC.
Tuesday July 12, 2011
- Liquid cultures of GFP1 (x2), IN1, IN2, GFP2 and PSB1C3 were innoculated with the appropriate antibiotic in the broth.
Wednesday July 13, 2011
- GFP1, IN1, IN2, GFP2 and PSB1C3 were minipreped to isolate plasmid DNA.
- Frozen stock of GFP1 made.
Thursday July 14, 2011
- GFP1, IN1, IN2, GFP2, PSB1C3 digested with EcoRI and PstI. GFP2 also digested with ndeI.
- Innoculation of liquid culture (GFP1, IN1, IN2, GFP2, PSB1C3).
Fridat July 15, 2011
- Gel extraction of GFP1, IN1, IN2, GFP2 and PSB1C3. However, the results were not as anticipated.
- Miniprep of cultures innoculated yesterday.
Monday July 18, 2011
- Cultures were miniprepped, however, GFP1 did not have a sufficient concentration to undergo digestion.
- Proceeded with digestion for GFP2 (ndeI), IN1, IN2 and PSBIC3 with EcoRI and PstI.
- Innoculation of GFP1 (x4)
Tuesday July 19, 2011
- Miniprep and digestion of GFP1.
- Gel extraction of each digestion (PSB1C3(x2), GFP1, IN1, IN2, GFP2).
Wednesday July 20, 2011
- Lox resuspended and digested.
- Ligation of GFP2, IN1, IN2 and Lox into PSB1C3.
Thursday July 21, 2011
- Transformation of GFP2, IN1, IN2 and Lox. Each was plated on cm containing media and grown overnight.
Friday July 22, 2011
- All negative plates did not produce colonies.
- Growth was good on all positive plates except for IN1, which only produced two main colonies.
Monday July 25, 2011
- Gel extraction IN1, IN2 and GFP1, however, GFP1 failed.
- Ligation of IN1 and IN2 into PSB1C3.
Tuesday July 26, 2011
- Innoculation of GFP2 into cm containing LB broth tube and lox into amp containing LB broth tube.
- Gel extraction did not work (likely a problem with digestion or transformation)
- Innoculation x5 of GFP1.
Wednesday July 27, 2011
- Frozen stock of GFP2 and lox made
- Miniprep of GFP1 x4 replicates
- Gel extraction of IN1 and IN2 resulted in improved concentrations
- Transformation
