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Team:Glasgow/LOVresults
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<h3>Aims</h3> | <h3>Aims</h3> | ||
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<br>1)Perform restriction digest to ensure we have LOV2 domain. | <br>1)Perform restriction digest to ensure we have LOV2 domain. | ||
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| - | 4)Streaked LOV2 containing Top10 onto 1mM IPTG and Ampicillan agar plates. Kept in dark for 3 hours at 30 degrees. | + | 4)Streaked LOV2 containing Top10 from glycerol stocks onto 1mM IPTG and Ampicillan agar plates. Kept in dark for 3 hours at 30 degrees. Used Blak Ray lamp, and transilluminator on plates to visualise any fluorescence. No fluorescence was seen. |
| + | Made up liquid cultures of Top10 + LOV2 in 2ml of LB + 2 microlitres Ampicillan. Liquid cultures grown for 4 hours and then 0.5 ml plated onto 1mM IPTG and Ampicillan plates. These were then wrapped in tinfoil and allowed to grow in the 30 degree incubator for 24 hours. | ||
<br>1) | <br>1) | ||
Revision as of 14:53, 21 September 2011
LOV2 Results
Aims
1)Perform restriction digest to ensure we have LOV2 domain. 2) Transform LOV2 domain into Top 10 3) Make glycerol stocks of LOV2 containing Top 10
2) Design site directed mutagenesis primers to get rid of illegal pst1 site
3)To perform site directed mutagenesis on LOV2
4)Make LOV2 fluoresce under UV light.
5)Ligate LOV2 into sumbission vector
6)Submit LOV 2
Methods
4)Streaked LOV2 containing Top10 from glycerol stocks onto 1mM IPTG and Ampicillan agar plates. Kept in dark for 3 hours at 30 degrees. Used Blak Ray lamp, and transilluminator on plates to visualise any fluorescence. No fluorescence was seen. Made up liquid cultures of Top10 + LOV2 in 2ml of LB + 2 microlitres Ampicillan. Liquid cultures grown for 4 hours and then 0.5 ml plated onto 1mM IPTG and Ampicillan plates. These were then wrapped in tinfoil and allowed to grow in the 30 degree incubator for 24 hours.1)
iLOV Results
Aims
1) Design iLOV construct to be synthesised
2) Get iLOV synthesised
3)Transform iLOV cells and make fluoresce
4)Ligate iLOV into sybmission vector
5)Submit iLOV
