Team:Tianjin/Notebook

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<p class="menu_head">Notebook Contents</p>
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<a href="https://2012.igem.org/Team:Tianjin/Notebook/Modeling">Modeling Notebook</a>
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<a href="https://2012.igem.org/Team:Tianjin/Notebook/HumanPractice">Human Practice Notebook</a>
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<p class="menu_head">July, 2012</p>
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<div id="text-content">
<center><span style="font-size:46px;font-family:Cambria;margin-top:10px;line-height:80%">Notes of Experiments</span></center>
<center><span style="font-size:46px;font-family:Cambria;margin-top:10px;line-height:80%">Notes of Experiments</span></center>
==July 6th, 2012==
==July 6th, 2012==
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##3M KOAc / 2M HOAc
##3M KOAc / 2M HOAc
#Experiment technologies training, such as making gel, gel electrophoresis.
#Experiment technologies training, such as making gel, gel electrophoresis.
-
[[File:TJU2012-Note-gel-1.png|center|500px|gel 1]]
+
[[File:TJU2012-Note-gel-1.png|center|350px|gel 1]]
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[[File:TJU2012-Note-gel-2.png|center|500px|gel 2]]
+
 
 +
 
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[[File:TJU2012-Note-gel-2.png|center|350px|gel 2]]
==July 11th, 2012==
==July 11th, 2012==
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##PCR principle and operation
##PCR principle and operation
##Gel Extraction
##Gel Extraction
-
[[File:TJU2012-Note-gel-3.png|center|500px|gel 3]]
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[[File:TJU2012-Note-gel-3.png|center|350px|gel 3]]
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[[File:TJU2012-Note-gel-4.png|center|500px|gel 4]]
+
 
 +
 
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[[File:TJU2012-Note-gel-4.png|center|350px|gel 4]]
==July 13th, 2012==
==July 13th, 2012==
1. Received the oligo and began to mutate 16S rrnB operator.
1. Received the oligo and began to mutate 16S rrnB operator.
-
[[File:TJU2012-Note-gen-1.png|center|500px|Gene1]]
+
[[File:TJU2012-Note-gen-1.png|center|350px|Gene1]]
2. Mutated RBS of RFP and checked through gel electrophoresis.
2. Mutated RBS of RFP and checked through gel electrophoresis.
-
[[File:TJU2012-Note-gen-2.png|center|500px|Gene2]]
+
[[File:TJU2012-Note-gen-2.png|center|250px|Gene2]]
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[[File:TJU2012-Note-gel-5.png|thumb|center|500px|~1800bp, constant with anticipation]]
+
[[File:TJU2012-Note-gel-5.png|thumb|center|250px|~1800bp, constant with anticipation]]
==July 14th, 2012==
==July 14th, 2012==
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==July 15th, 2012==
==July 15th, 2012==
#Extract plasmid of cells inculcated after one day.
#Extract plasmid of cells inculcated after one day.
-
#Enzyme test of the extracted plasmid.[[File:TJU2012-Note-gel-6.png|center|500px|Gel 6]]
+
#Enzyme test of the extracted plasmid.[[File:TJU2012-Note-gel-6.png|center|200px|Gel 6]]
#Inclucated the right cells in Liquid LB and added Ala partly.
#Inclucated the right cells in Liquid LB and added Ala partly.
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Analyse the results.
Analyse the results.
[[File:TJU2012-Note-form-1.png|center|500px|Form 1]]
[[File:TJU2012-Note-form-1.png|center|500px|Form 1]]
-
[[File:TJU2012-Note-fig-2.png|center|500px|Fig 2]]
+
[[File:TJU2012-Note-fig-2.png|center|300px|Fig 2]]
==July 24th, 2012==
==July 24th, 2012==
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==Aug. 2nd, 2012==
==Aug. 2nd, 2012==
-
#mutated the RBS of the Amp resistance gene.[[File:TJU2012-Note-gen-4.png|center|500px|Gene 4]]
+
#mutated the RBS of the Amp resistance gene.[[File:TJU2012-Note-gen-4.png|center|250px|Gene 4]]
#linked genes and transformed them into E. coli.
#linked genes and transformed them into E. coli.
==Aug. 4th, 2012==
==Aug. 4th, 2012==
Results of plates
Results of plates
-
[[File:TJU2012-Note-fig-3.png|center|500px|fig 3]]
+
[[File:TJU2012-Note-fig-3.png|center|300px|fig 3]]
[[File:TJU2012-Note-form-4.png|center|500px|form 4]]
[[File:TJU2012-Note-form-4.png|center|500px|form 4]]
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==Aug. 11th, 2012==
==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.
Verified part 1 and 2. Synthesized the two small parts and linked through overlap PCR. Verified through ligase digestion and then gel electrophoresis.
-
[[File:TJU2012-Note-fig-4.png|center|500px|fig 4]]
+
[[File:TJU2012-Note-fig-4.png|center|600px|fig 4]]
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[[File:TJU2012-Note-fig-5.png|center|500px|fig 5]]
[[File:TJU2012-Note-fig-5.png|center|500px|fig 5]]
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Verify part 3, 4 and 5. Verified through ligase digestion and then gel electrophoresis.
Verify part 3, 4 and 5. Verified through ligase digestion and then gel electrophoresis.
[[File:TJU2012-Note-fig-6.png|center|500px|fig 6]]
[[File:TJU2012-Note-fig-6.png|center|500px|fig 6]]
 +
 +
[[File:TJU2012-Note-fig-7.png|center|500px|fig 7]]
[[File:TJU2012-Note-fig-7.png|center|500px|fig 7]]
 +
 +
[[File:TJU2012-Note-fig-8.png|center|500px|fig 8]]
[[File:TJU2012-Note-fig-8.png|center|500px|fig 8]]
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==Aug. 18nd, 2012==
+
==Aug. 18th, 2012==
Transformed all the five parts into Yeast for Assembler in Yeast.
Transformed all the five parts into Yeast for Assembler in Yeast.
==Aug. 20th, 2012==
==Aug. 20th, 2012==
-
#Results of the transformation.[[File:TJU2012-Note-fig-9.png|center|500px|fig 9]]
+
#Results of the transformation.[[File:TJU2012-Note-fig-9.png|center|300px|fig 9]]
#Extract plasmids from the Yeast.
#Extract plasmids from the Yeast.
-
==Aug. 22th, 2012==
+
==Aug. 22nd, 2012==
-
#Transformed the plasmid from the Yeast into cells.[[File:TJU2012-Note-fig-10.png|thumb|center|500px|There are purple spots on the plates]]
+
#Transformed the plasmid from the Yeast into cells.[[File:TJU2012-Note-fig-10.png|thumb|center|300px|There are purple spots on the plates]]
-
#Extracted plasmid and checked through gel electrophoresis.[[File:TJU2012-Note-gel-7.png|center|500px|gel 7]]
+
#Extracted plasmid and checked through gel electrophoresis.[[File:TJU2012-Note-gel-7.png|center|300px|gel 7]]
 +
#Some other results of Assembler in Yeast.
 +
[[File:TJU2012-Note-fig-11.png|center|300px|fig 11]]
 +
 
 +
 
 +
[[File:TJU2012-Note-fig-12.png|center|300px|fig 12]]
 +
 
 +
 
 +
[[File:TJU2012-Note-fig-13.png|center|300px|fig 13]]
 +
 
 +
 
 +
[[File:TJU2012-Note-fig-14.png|center|300px|fig 14]]
==Aug. 25th, 2012==
==Aug. 25th, 2012==
-
Results of transformation.
+
#Began to build biobricks.[[File:Tianjin_BB2.PNG|center|500px|BB 2]]
 +
#Began to search articles on phages.
==Sept. 1st, 2012==
==Sept. 1st, 2012==
 +
Went to Peking University for exchanges.
 +
[[File:TJU2012-Note-fig-15.png|center|500px|fig 15]]
 +
 +
 +
[[File:TJU2012-Note-fig-16.png|center|500px|fig 16]]
 +
==Sept. 2nd, 2012==
==Sept. 2nd, 2012==
-
==Sept. 5st, 2012==
+
#Found out phi X174 and firstly proposed our ideas.
-
==Sept. 11st, 2012==
+
#The second sections of our biobricks.
-
==Sept. 18st, 2012==
+
[[File:Tianjin_BB1.PNG|center|500px|BB 1]]
 +
 
 +
==Sept. 5th, 2012==
 +
#Some experiments on phages.
 +
#Optimized our biobricks.
 +
#Designed primers needed for mutating Phi X174 and send orders.
 +
 
 +
==Sept. 11th, 2012==
 +
#Began to mutate gene G and E.
 +
#Optimized our biobricks.
 +
 
 +
==Sept. 18th, 2012==
 +
Began to sort out our materials for wiki and upload the website.
 +
[[File:new1.jpg|center|500px|fig 17]]
 +
our team in Hongkong
 +
 
 +
 
 +
[[File:new2.jpg|center|500px|fig 18]]
 +
our gold award
 +
 
 +
 
 +
[[File:new3.jpg|center|500px|fig 19]]
 +
presentation in MIT
{{:Team:Tianjin/footer}}
{{:Team:Tianjin/footer}}

Latest revision as of 05:16, 9 May 2013

Notebook Contents

Modeling Notebook
Human Practice Notebook

July, 2012

Sun
Mon
Tue
Wed
Thu
Fri
Sat
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

August, 2012

Sun
Mon
Tue
Wed
Thu
Fri
Sat
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

September, 2012

Sun
Mon
Tue
Wed
Thu
Fri
Sat
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

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


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


gel 4

July 13th, 2012

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

Gene1

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

Gene2
~1800bp, constant with anticipation

July 14th, 2012

  1. Transform two plasmids of we got yestday together into E.coli.
    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
  3. Inclucated the right cells in Liquid LB and added Ala partly.

July 20th, 2012

Analyse the results.

Form 1
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
  2. In the same method to construct three other systems.
fig 3
fig 4
fig 5

July 30th, 2012

  1. Learned Fluorescence spectrophotometer.
  2. measure the strength of GFP and RFP.
form 3

Aug. 2nd, 2012

  1. mutated the RBS of the Amp resistance gene.
    Gene 4
  2. linked genes and transformed them into E. coli.

Aug. 4th, 2012

Results of plates

fig 3
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


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


fig 7


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
  2. Extract plasmids from the Yeast.

Aug. 22nd, 2012

  1. Transformed the plasmid from the Yeast into cells.
    There are purple spots on the plates
  2. Extracted plasmid and checked through gel electrophoresis.
    gel 7
  3. Some other results of Assembler in Yeast.
fig 11


fig 12


fig 13


fig 14

Aug. 25th, 2012

  1. Began to build biobricks.
    BB 2
  2. Began to search articles on phages.

Sept. 1st, 2012

Went to Peking University for exchanges.

fig 15


fig 16

Sept. 2nd, 2012

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

our team in Hongkong


fig 18

our gold award


fig 19

presentation in MIT


Retrieved from "http://2012.igem.org/Team:Tianjin/Notebook"