iGEM 2012 — Team Tianjin

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# Team:Tianjin/Notebook

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Notebook Contents

[Modeling Notebook](https://2012.igem.org/Team%3ATianjin/Notebook/Modeling)

[Human Practice Notebook](https://2012.igem.org/Team%3ATianjin/Notebook/HumanPractice)

July, 2012

Sun

Mon

Tue

Wed

Thu

Fri

Sat

1

2

3

4

5

[6](#July_6th.2C_2012)

7

[8](#July_8th.2C_2012)

9

10

[11](#July_11th.2C_2012)

12

[13](#July_13th.2C_2012)

[14](#July_14th.2C_2012)

[15](#July_15th.2C_2012)

16

17

18

19

[20](#July_20th.2C_2012)

21

22

23

[24](#July_24th.2C_2012)

25

26

27

28

29

[30](#July_30th.2C_2012)

31

August, 2012

Sun

Mon

Tue

Wed

Thu

Fri

Sat

1

[2](#Aug._2nd.2C_2012)

3

[4](#Aug._4th.2C_2012)

5

6

7

[8](#Aug._8th.2C_2012)

9

10

[11](#Aug._11th.2C_2012)

[12](#Aug._12th.2C_2012)

13

[14](#Aug._14th.2C_2012)

15

16

17

[18](#Aug._18th.2C_2012)

19

[20](#Aug._20th.2C_2012)

21

[22](#Aug._22nd.2C_2012)

23

24

[25](#Aug._25th.2C_2012)

26

27

28

29

30

31

September, 2012

Sun

Mon

Tue

Wed

Thu

Fri

Sat

[1](#Sept._1st.2C_2012)

[2](#Sept._2nd.2C_2012)

3

4

[5](#Sept._5th.2C_2012)

6

7

8

9

10

[11](#Sept._11th.2C_2012)

12

13

14

15

16

17

[18](#Sept._18th.2C_2012)

19

20

21

22

23

24

25

26

27

28

29

30

Notes of Experiments

## July 6th, 2012

1. We discuss our all of the possible projects, and finally choose one. And then allocate works to specific member.
2. Do some pre-experiment and make some reagants.
   1. Liquid LB
      1. 10g tryptone
      2. 5g yeast extract
      3. 10g NaCl
   2. Solid LB
      1. 10g tryptone
      2. 5g yeast extract
      3. 10g NaCl
      4. 20g agar
3. Cleaned up the lab and laboratory instruments

## July 8th, 2012

1. Solutions for extracting plasmid.
   1. 50mM Glucose / 25mM Tris-Cl / 10mM EDTA，pH=8.0
   2. 0.2N NaOH / 1% SDS
   3. 3M KOAc / 2M HOAc
2. Experiment technologies training, such as making gel, gel electrophoresis.

[![gel 1](/wiki/images/1/14/TJU2012-Note-gel-1.png)](/File%3ATJU2012-Note-gel-1.png "gel 1")

[![gel 2](/wiki/images/5/5b/TJU2012-Note-gel-2.png)](/File%3ATJU2012-Note-gel-2.png "gel 2")

## July 11th, 2012

1. Designed primers and sent orders.
2. Optimized the project and allocated work.
3. Experiment technologies and knowledge training
   1. PCR principle and operation
   2. Gel Extraction

[![gel 3](/wiki/images/f/fb/TJU2012-Note-gel-3.png)](/File%3ATJU2012-Note-gel-3.png "gel 3")

[![gel 4](/wiki/images/3/35/TJU2012-Note-gel-4.png)](/File%3ATJU2012-Note-gel-4.png "gel 4")

## July 13th, 2012

1. Received the oligo and began to mutate 16S rrnB operator.

[![Gene1](/wiki/images/1/14/TJU2012-Note-gen-1.png)](/File%3ATJU2012-Note-gen-1.png "Gene1")

2. Mutated RBS of RFP and checked through gel electrophoresis.

[![Gene2](/wiki/images/6/62/TJU2012-Note-gen-2.png)](/File%3ATJU2012-Note-gen-2.png "Gene2")

[![](/wiki/images/2/2a/TJU2012-Note-gel-5.png)](/File%3ATJU2012-Note-gel-5.png)

[![](/wiki/skins/common/images/magnify-clip.png)](/File%3ATJU2012-Note-gel-5.png "Enlarge")

~1800bp, constant with anticipation

## July 14th, 2012

1. Transform two plasmids of we got yestday together into E.coli.

   [![Gene 3](/wiki/images/e/ec/TJU2012-Note-gen-3.png)](/File%3ATJU2012-Note-gen-3.png "Gene 3")
2. PCR verification of colonies on the LB plates.
3. Inculcated the right colonies in Liquid LB.

## July 15th, 2012

1. Extract plasmid of cells inculcated after one day.
2. Enzyme test of the extracted plasmid.

   [![Gel 6](/wiki/images/c/c4/TJU2012-Note-gel-6.png)](/File%3ATJU2012-Note-gel-6.png "Gel 6")
3. Inclucated the right cells in Liquid LB and added Ala partly.

## July 20th, 2012

Analyse the results.

[![Form 1](/wiki/images/4/4f/TJU2012-Note-form-1.png)](/File%3ATJU2012-Note-form-1.png "Form 1")

[![Fig 2](/wiki/images/9/90/TJU2012-Note-fig-2.png)](/File%3ATJU2012-Note-fig-2.png "Fig 2")

## July 24th, 2012

1. Linked the GRP gene and O-RBS RFP gene. Gel extract the previous product again, and ligate using T4 with the following gradient.

   [![form 2](/wiki/images/e/ea/TJU2012-Note-form-2.png)](/File%3ATJU2012-Note-form-2.png "form 2")
2. In the same method to construct three other systems.

[![fig 3](/wiki/images/6/6c/TJU2012-Mode-cal-fig-2.png)](/File%3ATJU2012-Mode-cal-fig-2.png "fig 3")

[![fig 4](/wiki/images/d/db/TJU2012-Mode-cal-fig-3.png)](/File%3ATJU2012-Mode-cal-fig-3.png "fig 4")

[![fig 5](/wiki/images/9/9a/TJU2012-Mode-cal-fig-4.png)](/File%3ATJU2012-Mode-cal-fig-4.png "fig 5")

## July 30th, 2012

1. Learned Fluorescence spectrophotometer.
2. measure the strength of GFP and RFP.

[![form 3](/wiki/images/1/13/TJU2012-Note-form-3.png)](/File%3ATJU2012-Note-form-3.png "form 3")

## Aug. 2nd, 2012

1. mutated the RBS of the Amp resistance gene.

   [![Gene 4](/wiki/images/c/c4/TJU2012-Note-gen-4.png)](/File%3ATJU2012-Note-gen-4.png "Gene 4")
2. linked genes and transformed them into E. coli.

## Aug. 4th, 2012

Results of plates

[![fig 3](/wiki/images/8/83/TJU2012-Note-fig-3.png)](/File%3ATJU2012-Note-fig-3.png "fig 3")

[![form 4](/wiki/images/5/59/TJU2012-Note-form-4.png)](/File%3ATJU2012-Note-form-4.png "form 4")

## Aug. 8th, 2012

Began to construct the five parts needed in Assembler in Yeast. Part 1 and part 2.

## Aug. 11th, 2012

Verified part 1 and 2. Synthesized the two small parts and linked through overlap PCR. Verified through ligase digestion and then gel electrophoresis.

[![fig 4](/wiki/images/0/04/TJU2012-Note-fig-4.png)](/File%3ATJU2012-Note-fig-4.png "fig 4")

[![fig 5](/wiki/images/4/4c/TJU2012-Note-fig-5.png)](/File%3ATJU2012-Note-fig-5.png "fig 5")

## Aug. 12th, 2012

Began to build part 3, 4 and 5. Synthesized the two small parts and linked through overlap PCR.

## Aug. 14th, 2012

Verify part 3, 4 and 5. Verified through ligase digestion and then gel electrophoresis.

[![fig 6](/wiki/images/5/59/TJU2012-Note-fig-6.png)](/File%3ATJU2012-Note-fig-6.png "fig 6")

[![fig 7](/wiki/images/0/00/TJU2012-Note-fig-7.png)](/File%3ATJU2012-Note-fig-7.png "fig 7")

[![fig 8](/wiki/images/9/96/TJU2012-Note-fig-8.png)](/File%3ATJU2012-Note-fig-8.png "fig 8")

## Aug. 18th, 2012

Transformed all the five parts into Yeast for Assembler in Yeast.

## Aug. 20th, 2012

1. Results of the transformation.

   [![fig 9](/wiki/images/d/d8/TJU2012-Note-fig-9.png)](/File%3ATJU2012-Note-fig-9.png "fig 9")
2. Extract plasmids from the Yeast.

## Aug. 22nd, 2012

1. Transformed the plasmid from the Yeast into cells.

   [![](/wiki/images/b/bc/TJU2012-Note-fig-10.png)](/File%3ATJU2012-Note-fig-10.png)

   [![](/wiki/skins/common/images/magnify-clip.png)](/File%3ATJU2012-Note-fig-10.png "Enlarge")

   There are purple spots on the plates
2. Extracted plasmid and checked through gel electrophoresis.

   [![gel 7](/wiki/images/a/a7/TJU2012-Note-gel-7.png)](/File%3ATJU2012-Note-gel-7.png "gel 7")
3. Some other results of Assembler in Yeast.

[![fig 11](/wiki/images/6/69/TJU2012-Note-fig-11.png)](/File%3ATJU2012-Note-fig-11.png "fig 11")

[![fig 12](/wiki/images/8/84/TJU2012-Note-fig-12.png)](/File%3ATJU2012-Note-fig-12.png "fig 12")

[![fig 13](/wiki/images/7/7f/TJU2012-Note-fig-13.png)](/File%3ATJU2012-Note-fig-13.png "fig 13")

[![fig 14](/wiki/images/1/14/TJU2012-Note-fig-14.png)](/File%3ATJU2012-Note-fig-14.png "fig 14")

## Aug. 25th, 2012

1. Began to build biobricks.

   [![BB 2](/wiki/images/5/52/Tianjin_BB2.PNG)](/File%3ATianjin_BB2.PNG "BB 2")
2. Began to search articles on phages.

## Sept. 1st, 2012

Went to Peking University for exchanges.

[![fig 15](/wiki/images/5/59/TJU2012-Note-fig-15.png)](/File%3ATJU2012-Note-fig-15.png "fig 15")

[![fig 16](/wiki/images/1/1b/TJU2012-Note-fig-16.png)](/File%3ATJU2012-Note-fig-16.png "fig 16")

## Sept. 2nd, 2012

1. Found out phi X174 and firstly proposed our ideas.
2. The second sections of our biobricks.

[![BB 1](/wiki/images/0/02/Tianjin_BB1.PNG)](/File%3ATianjin_BB1.PNG "BB 1")

## Sept. 5th, 2012

1. Some experiments on phages.
2. Optimized our biobricks.
3. Designed primers needed for mutating Phi X174 and send orders.

## Sept. 11th, 2012

1. Began to mutate gene G and E.
2. Optimized our biobricks.

## Sept. 18th, 2012

Began to sort out our materials for wiki and upload the website.

[![fig 17](/wiki/images/4/49/New1.jpg)](/File%3ANew1.jpg "fig 17")

our team in Hongkong

[![fig 18](/wiki/images/a/a8/New2.jpg)](/File%3ANew2.jpg "fig 18")

our gold award

[![fig 19](/wiki/images/4/4a/New3.jpg)](/File%3ANew3.jpg "fig 19")

presentation in MIT

E-mail：austinamens@gmail.com | Address:Building No.20, No.92 Weijin Road, Tianjin, China | Zip-code：300072

Copyright 2012 © Tianjin University iGEM Team

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