Team:Newcastle/Labwork/4 August 2009
From 2009.igem.org
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===Preparation=== | ===Preparation=== | ||
- | Items needed: | + | ;Items needed: |
* Lysozyme (not available, ordered) | * Lysozyme (not available, ordered) | ||
* 10mM (0.01M) Potassium phosphate (Crystal form available, ordered) | * 10mM (0.01M) Potassium phosphate (Crystal form available, ordered) | ||
Line 28: | Line 28: | ||
- | 0.01M Potassium Phosphate Solution: | + | ;0.01M Potassium Phosphate Solution: |
+ | MW * Desired Volume (L) * Desired Molarity (M) | ||
+ | 174.18 * 0.1L * 0.01M = 0.174g | ||
+ | |||
+ | Therefore, 0.174g of Potassium Phosphate is mixed with 0.1L of DI water to make 0.1L of 0.01M Potassium Phosphate | ||
+ | |||
+ | |||
+ | ;0.05M KCl Solution: | ||
+ | 1M (Stock Solution) / 0.05M (Desired Molarity) = 20 | ||
+ | Therefore, in order to obtain a solution of 0.05M, the stock solution needs to be diluted 20 times. | ||
+ | |||
+ | Desired volume = 0.1L | ||
+ | 100ml/20=5ml of KCl, and DI water would make up 95ml, producing 100ml of 0.05M KCl. | ||
+ | |||
+ | |||
Revision as of 17:49, 4 August 2009
Lab Session 04/08/2009
Sporulation Tuning Team
Prof. Anne Moir from Sheffield University kindly sent us the spores of the genes cwlD and sleB/cwlJ
- HC401 (FrpC2, cwlJ:: Kan, sleB-lacZ Ery R)
- AM1877 (cwlD,Cm r, trp C2)
In the protocol given to us by Prof. Moir, there are two methods (Methods A and B), which can be used to recover the cwlD spores. However, neither give complete restoration of germination.
[Attach Prof. Moir's protocols]
Referring to Prof. Moir's protocols, Method A is a fast method which results in partial germination of approximately 0.1%. Method B on the other hand, is a slow method which involves stripping of the spore coat layers for improved germination of approximately 10% recovery. Due to the lack of time, the group decided to go with Method A.
Preparation
- Items needed
- Lysozyme (not available, ordered)
- 10mM (0.01M) Potassium phosphate (Crystal form available, ordered)
- 50mM (0.05M) KCl (Stock solution of 1M available)
- 1mM (0.001M) MgCl2 (Stock solution of 0.5M available)
- L-alanine (not available, ordered)
- 0.01M Potassium Phosphate Solution
MW * Desired Volume (L) * Desired Molarity (M) 174.18 * 0.1L * 0.01M = 0.174g
Therefore, 0.174g of Potassium Phosphate is mixed with 0.1L of DI water to make 0.1L of 0.01M Potassium Phosphate
- 0.05M KCl Solution
1M (Stock Solution) / 0.05M (Desired Molarity) = 20 Therefore, in order to obtain a solution of 0.05M, the stock solution needs to be diluted 20 times.
Desired volume = 0.1L 100ml/20=5ml of KCl, and DI water would make up 95ml, producing 100ml of 0.05M KCl.
News
Events
- 20 – 21 June 2009 - Europe workshop (London)
- 23 – 24 June 2009 - UK iGEM meetup (Edinburgh)
- 23 October Practice Presentation (Newcastle)
- 23 October T-shirts are ready
- 27 October Practice Presentation (Sunderland)
- 27 October Poster is ready
- 30 October – 2 November 2009 - Jamboree (Boston)
Social Net
- Newcastle iGEM Twitter
- [http://www.facebook.com/home.php#/group.php?gid=131709337641 Newcastle on Facebook]
- [http://www.youtube.com/user/newcastle2009igem Newcastle Youtube Channel]