Team:Peking S/lab/notebook/dln

From 2011.igem.org

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== '''summary''' ==
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== '''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.
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.
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===7.3===
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===7.3-7.9===
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Designed the structure of cell A, which was consisted of two plasmids:
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===7.4===
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PBad+RBS+RFP-tag+RBS+lasI+terminator 
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===7.5===
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PSal+RBS+ccdB+terminator
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===7.6===
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===7.10-7.16===
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pre-experiment for choosing the proper RBS(B0033) for ccdB
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===7.8===
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===7.9===
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===7.10===
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===7.12===
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===7.13===
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===7.14===
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===7.15===
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===7.16===
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===7.18===
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===7.19===
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===7.20===
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===7.21===
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===7.22===
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===7.23===
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chose the template mCherry(J06504 in partregistry) for PCR to add tag in the tail of mCherry
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===7.29===
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===7.18-7.24===
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PCR to add tag in the tail of mCherry
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===7.30===
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Did the ligation of RBS(B0033) and ccdB(K145151)
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===7.31===
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===7.29-7.31===
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The RBS+ccdB wasn't sequenced correct.Did the ligation again.
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The primer for adding tag on the mCherry wasn't correct.Resigned.
==August==
==August==
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===8.1===
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===8.1-8.7===
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The RBS+ccdB was sequenced correct.
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===8.2===
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Did the ligation of RBS+ccdB and terminator(B0015)
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===8.4===
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===8.8-8.14===
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PCR for adding tag on the mCherry.
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===8.5===
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The RBS+ccdB+terminator was sequenced correct.
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===8.6===
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PCR to acquire the promotor PSal.
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===8.7===
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===8.15-8.21===
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Did the ligation of PBad and mCherry-tag.
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===8.8===
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Did the ligation of PSal and RBS+ccdB+terminator.
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===8.9===
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===8.22-8.28===
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The PBad+mCherry-tag was sequenced correct.
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===8.10===
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The PSal+RBS+ccdB+terminator was sequenced correct.
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Did the ligation of PBad+mCherry-tag and lasI.
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===8.11===
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===8.30-8.31===
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The PSal+mCherry-tag+lasI was sequenced correct.
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===8.12===
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Prepared the backbone pSB1A3, pSB4K5, pSB1C3 to be digested.
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===8.13===
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===8.14===
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===8.15===
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===8.16===
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===8.17===
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===8.18===
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===8.19===
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===8.20===
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===8.21===
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===8.22===
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===8.23===
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===8.25===
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===8.26===
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===8.27===
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===8.30===
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===8.31===
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==September==
==September==
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===9.1===
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===9.1-9.7===
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Changed the backbone:
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===9.2===
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PBad+mCherry-tag+lasI:pSB1AK3->pSB1A3
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===9.3===
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PSal+RBS+ccdB+terminator:pSB1AK3->pSB4K5
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===9.6===
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RBS+ccdB+terminator:pSB1AK3->pSB1C3
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===9.7===
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===9.8-9.14===  
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Made the plan for human practice.
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===9.8===  
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===9.15-9.21===
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Made the survey of human practice
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===9.9===
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Sent out the questionaires to students in high school and university
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===9.12===
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===9.22-9.28===
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Literature review for synthetic biology and genetically engineered organism
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===9.13===
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===9.29-9.30===
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Wrote the article for human practice.
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===9.15===
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===9.16===
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===9.17===
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===9.21===
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===9.22===
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===9.23===
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===9.24===
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===9.25===
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===9.26===
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===9.27===
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===9.28===
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===9.29===
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===9.30===
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==October==
==October==
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===10.1===
===10.1===
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Wrote the notebook on the wiki.
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===10.3===
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modified the article of human practice.
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===10.4===
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===10.5===
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===10.7===
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===10.8===
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===10.9===
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===10.10===
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===10.11===
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===10.12===
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===10.13===
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===10.15===
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===10.16-10.21===
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===10.21-10.25===
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Latest revision as of 21:59, 5 October 2011


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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]

9.1-9.7

Changed the backbone:

PBad+mCherry-tag+lasI:pSB1AK3->pSB1A3

PSal+RBS+ccdB+terminator:pSB1AK3->pSB4K5

RBS+ccdB+terminator:pSB1AK3->pSB1C3

9.8-9.14

Made the plan for human practice.

9.15-9.21

Made the survey of human practice

Sent out the questionaires to students in high school and university

9.22-9.28

Literature review for synthetic biology and genetically engineered organism

9.29-9.30

Wrote the article for human practice.

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

Wrote the notebook on the wiki.

modified the article of human practice.











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