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Team:Peking S/lab/notebook/sr
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| - | == ''' | + | == '''Summary''' == |
| - | + | In this summer, I mainly focused on the construction of the microbial population density balancer. In addition, I also carried out the assay for orthogonality of positive regulating signaling molecules. | |
=='''Contents'''== | =='''Contents'''== | ||
| Line 85: | Line 85: | ||
===7.7=== | ===7.7=== | ||
| - | + | ||
| + | '''ABBREVIATIONS: | ||
| + | |||
| + | '''Restriction endonuleases: | ||
| + | |||
| + | '''E=EcoRI X=XbaI S=SpeI P=PstI''''' | ||
| + | ''' | ||
| + | |||
| + | |||
| + | |||
| + | 17:30 Digestion of B0034(1-2M) with S&P and 4-12I with X&P started. | ||
| + | Fragment lengths: B0034 around 2000bps | ||
| + | 4-12I 756 bps | ||
| + | PCR of terminator-less version of Ptet-rhlI and lacI(I732011)-LVA | ||
| + | |||
| + | Electroperesis: | ||
| + | Gel 1 Well 2-4: Marker_4-12I_1-2M | ||
| + | Gel 2 Well 2-5: Marker_rhlI_rhlI_rhlI | ||
| + | Gel 3 Well 1-6: Marker_lacI_lacI_ _ _lacI_lacI | ||
| + | All checked right. | ||
| + | |||
| + | Gel extraction of the above DNA fragments. | ||
| + | |||
| + | Ligation of B0034(SP digested)+4-12I(XP digested). | ||
| + | |||
===7.8=== | ===7.8=== | ||
| + | |||
| + | 12:30 Transformation of the ligation product(1-2M+4-12I) finished. | ||
| + | 12:30 Gel extraction of PCR product (LacI-LVA) finished. | ||
| + | |||
| + | 17:30 PCR for lacI-tag from I732100 started. | ||
| + | |||
| + | 19:30 Transformation of 2-10G finished. | ||
| + | |||
| + | 19:30 PCR failed. Repeated with the same system &condition and got the right string. | ||
===7.9=== | ===7.9=== | ||
| + | 0 :19 Checked plates. | ||
| + | |||
| + | 0 :19 Gel extraction & PCR of lacI-LVA-suffix. | ||
| + | |||
| + | 14:00 Digestion of 1-1G and lacI PCR product(second round) with X&P. | ||
| + | |||
| + | 14:00 Colony pick of 1-1G plates. | ||
| + | |||
| + | 18:30 Electrophoresis for digestion products(1-1G XP fragment;lacI PCR product digested with X&P). | ||
| + | |||
| + | 23:45 Ligation of 1-1G and lacI-suffix fragments started. | ||
===7.10=== | ===7.10=== | ||
| + | 00:05 Digestion of Ptet-rhlI PCR products (1 and 2) started. | ||
| + | 00:05 Digestions: 2-10G(EX) 1-2M+4-12I(RBS+gfp-LVA)(1,2,3，ES) 1-23L（EX&ES) started. | ||
===7.12=== | ===7.12=== | ||
Latest revision as of 05:20, 5 October 2011
Template:Https://2011.igem.org/Team:Peking S/bannerhidden
- Rui Sun's Notebook
Summary
In this summer, I mainly focused on the construction of the microbial population density balancer. In addition, I also carried out the assay for orthogonality of positive regulating signaling molecules.
Contents
July
| Mon | Tue | Wed | Thu | Fri | Sat | Sun |
| - | - | - | - | 1 | 2 | 3 |
| 4 | 5 | 6 | 7 | 8 | 9 | 10 |
| 11 | 12 | 13 | 14 | 15 | 16 | 17 |
| 18 | 19 | 20 | 21 | 22 | 23 | 24 |
| 25 | 26 | 27 | 28 | 29 | 30 | 31 |
| - | - | - | - | - | - | - |
[TOP]
7.7
ABBREVIATIONS:
Restriction endonuleases:
E=EcoRI X=XbaI S=SpeI P=PstI
17:30 Digestion of B0034(1-2M) with S&P and 4-12I with X&P started.
Fragment lengths: B0034 around 2000bps
4-12I 756 bps
PCR of terminator-less version of Ptet-rhlI and lacI(I732011)-LVA
Electroperesis:
Gel 1 Well 2-4: Marker_4-12I_1-2M
Gel 2 Well 2-5: Marker_rhlI_rhlI_rhlI
Gel 3 Well 1-6: Marker_lacI_lacI_ _ _lacI_lacI
All checked right.
Gel extraction of the above DNA fragments.
Ligation of B0034(SP digested)+4-12I(XP digested).
7.8
12:30 Transformation of the ligation product(1-2M+4-12I) finished. 12:30 Gel extraction of PCR product (LacI-LVA) finished.
17:30 PCR for lacI-tag from I732100 started.
19:30 Transformation of 2-10G finished.
19:30 PCR failed. Repeated with the same system &condition and got the right string.
7.9
0 :19 Checked plates.
0 :19 Gel extraction & PCR of lacI-LVA-suffix.
14:00 Digestion of 1-1G and lacI PCR product(second round) with X&P.
14:00 Colony pick of 1-1G plates.
18:30 Electrophoresis for digestion products(1-1G XP fragment;lacI PCR product digested with X&P).
23:45 Ligation of 1-1G and lacI-suffix fragments started.
7.10
00:05 Digestion of Ptet-rhlI PCR products (1 and 2) started. 00:05 Digestions: 2-10G(EX) 1-2M+4-12I(RBS+gfp-LVA)(1,2,3，ES) 1-23L（EX&ES) started.
7.12
7.13
7.14
7.15
7.16
7.17
7.18
7.19
7.20
7.21
7.22
7.23
7.24
7.25
7.26
7.27
7.28
7.29
7.30
7.31
August
| Mon | Tue | Wed | Thu | Fri | Sat | Sun |
| 1 | 2 | 3 | 4 | 5 | 6 | 7 |
| 8 | 9 | 10 | 11 | 12 | 13 | 14 |
| 15 | 16 | 17 | 18 | 19 | 20 | 21 |
| 22 | 23 | 24 | 25 | 26 | 27 | 28 |
| 29 | 30 | 31 | - | - | - | - |
[TOP]
8.1
8.2
8.4
8.5
8.6
8.7
8.8
8.9
8.10
8.11
8.12
8.13
8.14
8.15
8.16
8.17
8.18
8.19
8.20
8.21
8.22
8.23
8.25
8.26
8.27
8.30
8.31
September
| Mon | Tue | Wed | Thu | Fri | Sat | Sun |
| - | - | 1 | 2 | 3 | 4 | |
| 5 | 6 | 7 | 8 | 9 | 10 | 11 |
| 12 | 13 | 14 | 15 | 16 | 17 | 18 |
| 19 | 20 | 21 | 22 | 23 | 24 | 25 |
| 26 | 27 | 28 | 29 | 30 | - | -- |
[TOP]
9.1
9.2
9.3
9.6
9.7
9.8
9.9
9.12
9.13
9.14
9.15
9.16
9.17
9.21
9.22
9.23
9.24
9.25
9.26
9.27
9.28
9.29
9.30
October
| Mon | Tue | Wed | Thu | Fri | Sat | Sun |
| - | - | - | - | 1 | 2 | 3 |
| 4 | 5 | 6 | 7 | 8 | 9 | 10 |
| 11 | 12 | 13 | 14 | 15 | 16 | 17 |
| 18 | 19 | 20 | 21 | 22 | 23 | 24 |
| 25 | - | - | - | - | - | - |
[TOP]
10.1
10.3
10.4
10.5
10.7
10.8
10.9
10.10
10.11
10.12
10.13
10.15
10.16-10.21
10.21-10.25
