Team:UCL/Labbook/Week16
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Lab Weeks
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Week 16
Bacterial Labs
Results of the inoculations of transformations of 13/09 in 4xcmp: all showed growth apart from falcons 5.3, 5.2, 1.14 and 1.2. Glycerol stocks of the rest of 19 inoculations were made.
Minipreps of the above inoculations were made only for 1.3, 1.7, 1.9, 1.10, 1.11, 1.15, 1.20 and 5.1 due to lack of chromatographic columns.
Nanodrop result of the above minipreps showed concentration values below 16.0 ng/ul. Analytical digest of miniprep samples 1-9 (prepared the day before) as well as of 5.1 and 1.15 (which showed concentrations of about 15 ng/ul) with E and P
Components | Volume (ul) |
---|---|
DNA | 5 |
EcoR1 | 1 |
Pst1 | 1 |
Buffer 3 | 1 |
BSA | 0.5 |
dH2O | 1.5 |
Total | 10 |
INSERT IMAGE OF GEL 16/09 ZEO BB CANDIDATES DIGESTED WITH E&P FROM MINIPREPS OF COLONIES TRANSFORMED ON 13/09
Prep digest of miniprep of J632014 with E and P in order to keep up the pSB1C3 stocks
Components | Cut | Uncut | Control - pSecTag2A |
---|---|---|---|
J632014 DNA | 25 | 5 | 10 |
EocR1 | 2 | 0 | 0 |
Pst1 | 2 | 0 | 0 |
Buffer 3 | 4 | 0 | 0 |
BSA | 0.5 | 0 | 0 |
dH2O | 6.5 | 5 | 0 |
Total | 40 | 10 | 10 |
These were incubated for 2 hours at 37O C. Casey ran a gel of the following samples in the next order: Zeo BB: 26D, 26u; 28D, 28u; AuxD, AuxU INSERT IMAGE OF GEL HERE PLS
Gel1: E+P double digest and P, single digest (Auxin bb): 3, 12, 13, 15. Gel2: D digest 26, 28 and Auxin bb. Minipreps: 26, 28, 3, 12, 13, 15, 21 (sample of which DNA is waiting to be eluted)
Tuesday 17th September
Miniprep of the 16 tubes of inoculations of MMP9 as well as tube 21 of which DNA had to be eluted.
Nanodrop results of the minipreps
Tube no. | ng/ul | 260/280 |
---|---|---|
1 | 20.6 | 1.9 |
2 | 26.9 | 1.64 |
4 | 22.0 | 3.2 |
5 | 4.8 | 2.16 |
6 | 22.7 | 2.99 |
7 | 16.4 | 2.41 |
8 | 16.9 | 2.01 |
9 | 7.1 | 1.49 |
10 | 27.3 | 1.79 |
11 | 16.0 | 1.72 |
12 | 22.2 | 1.41 |
13 | 11.0 | 1.73 |
14 | 12.0 | 1.6 |
15 | 18.4 | 1.57 |
17 | 45.1 | 1.62 |
18 | 9.2 | 1.72 |
21 (kc) | 21.3 | 1.81 |