Team:Newcastle/7 September 2010

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(→First transformation of B. subtilis 168 containing pMutin4 with pGFPrrnB containing yneA)
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=First transformation of ''B. subtilis'' 168 containing pMutin4 with pGFPrrnB containing ''yneA''=
 
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===Aim===
 
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The aim of the experiment is to perform insert the plasmid pGFP-rrnB containing ''yneA'' which have been ligated eariler into the chromosome of ''Bacillus subtilis'' 168. ''B. subtilis'' containing the intergated vector will be resistance to both the antibiotics chloramphenicol and streptomycin, therefore those that have successful intergated will be selected with agar plates that contain both the antibiotics. The second step will be to identify those colones that have the plasmid intergated at the corerct position in the chromosome, which is the amylase gene locus. Thus those that have intergrated at the wrong position will not be able to break down starch, which can be tested with the iodine test.
 
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===Materials and Protocol===
 
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Please refer to: [[Team:Newcastle/Transformation of B. subtilis| Transformation of ''B. subtilis'']]
 
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Note: Overnight culture of ''B. subtilis'' 168 in MM competence medium was done the day before and the iodine test was performed the day after.
 
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===Result===
 
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The transformation was unsuccessful.
 
=Subtilin Immunity BioBrick=
=Subtilin Immunity BioBrick=

Revision as of 14:28, 27 October 2010

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Contents

Subtilin Immunity BioBrick

Aims

The aim of this experiment is to perform minipreps of the 16 overnight cultures that were left overnight from yesterday for the Subtilin Immunity BioBrick.

Methods

The Qiagen Miniprep protocol was used for each of the 16 cultures, followed by the NanoDrop Spectrophotometer protocol.

Results, Discussion and Conclusion

The nanodrop spectrophotometer results for the 16 minipreps for Subtilin Immunity are below (units: ng/ml):

Tube Nanodrop Value (in ng/ml)
1 76.5
2 61.5
3 84.6
4 19.5
5 89.9
6 87.6
7 6.3
8 69.5
9 81.3
10 74.4
11 68.5
12 70.2
13 83.5
14 77.6
15 73.3
16 63.0


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