"
Team:Peking S/lab/notebook/dln
From 2011.igem.org
Template:Https://2011.igem.org/Team:Peking S/bannerhidden
Linna Deng's Notebook
summary
After designed the gene circuit in cell A, I constructed a duoble-plasmids stucture containing the expression of signal molecules and death protein. One of the plasmid is consisted of an PBad promoter without AraC, the gene expressing mCherry, and lasI protein generator. the other is made up with promoter for salicylic acid and ccdB gene which is a control of cell death. I used PCR to confirm whether the plasmids containing the genes were in correct size.
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]
7.3-7.9
Designed the structure of cell A, which was consisted of two plasmids:
PBad+RBS+RFP-tag+RBS+lasI+terminator
PSal+RBS+ccdB+terminator
7.10-7.16
pre-experiment for choosing the proper RBS(B0033) for ccdB
chose the template mCherry(J06504 in partregistry) for PCR to add tag in the tail of mCherry
7.18-7.24
PCR to add tag in the tail of mCherry
Did the ligation of RBS(B0033) and ccdB(K145151)
7.29-7.31
The RBS+ccdB wasn't sequenced correct.Did the ligation again.
The primer for adding tag on the mCherry wasn't correct.Resigned.
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]
8.1-8.7
The RBS+ccdB was sequenced correct.
Did the ligation of RBS+ccdB and terminator(B0015)
8.8-8.14
PCR for adding tag on the mCherry.
The RBS+ccdB+terminator was sequenced correct.
PCR to acquire the promotor PSal.
8.15-8.21
Did the ligation of PBad and mCherry-tag.
Did the ligation of PSal and RBS+ccdB+terminator.
8.22-8.28
The PBad+mCherry-tag was sequenced correct.
The PSal+RBS+ccdB+terminator was sequenced correct.
Did the ligation of PBad+mCherry-tag and lasI.
8.30-8.31
The PSal+mCherry-tag+lasI was sequenced correct.
Prepared the backbone pSB1A3, pSB4K5, pSB1C3 to be digested.
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
