Team:Kyoto/CiC/Notebook/0928-1002

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-
==0914-0918 : what to do. what to do. what to do. what to do. ==
+
==0928-1004 : constructing sig-GFP generator and sig-EGFP generator(HeLa) ==
 +
 
{{:Team:Kyoto/Pad_start_dates}}
{{:Team:Kyoto/Pad_start_dates}}
-
===Monday, 14 August===
+
===Monday, 28 September===
====To Do====
====To Do====
-
*p-2-(1)(4)(7)(8)(10)
+
*sig-EGFP generator(HeLa)
-
**Miniprep
+
**Restriction Enzyme Digestion(Hind3)
-
**Restriction Enzyme Digestion
+
-
*p-2-(1)(4)  E.X
+
-
**PCR purification kit
+
-
*p-2-(1)(7)(8)(10)  E.S
+
-
**Gel extraction
+
-
*4-(1)(2)(3)
+
-
**Colony PCR
+
-
**Insert check
+
-
 
+
====Results====
====Results====
-
=====p-2-(1)(4)(7)(8)(10)=====
+
They were unexpectedly digested by Hind3,the Restriction Enzyme which would not work if the signal sequence were inserted in the vector correctly.
-
=====p-2-(1)(7)(8)(10)  E.S=====
+
-
*Gel extraction kit
+
-
[[Image:Kyoto_0914_PCR.png|250px]]
+
-
{| class="table"
+
-
|+Miniprep product conc.
+
-
!NO.!!Sample name!!conc./(ng/ul)
+
-
|-
+
-
|1||p-2-(1)||9.3
+
-
|-
+
-
|2||p-2-(7)||25.9
+
-
|-
+
-
|3||p-2-(8)||8.6
+
-
|-
+
-
|4||p-2-(10)||7.5
+
-
|}
+
-
 
+
-
=====4-(1)(2)(3)=====
+
-
[[Image:Kyoto_0914_PCR_2.png|250px]]
+
-
*This result indicates that (1)a, (2)b and (3)a were inserted properly.
+
-
 
+
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-
===Tuesday, 15 September===
 
-
====To Do====
 
-
'''Further to 14 Sept.'''
 
-
 
-
*p-2-(1)(4)(7)(8)(10)
 
-
**Miniprep
 
-
**Restriction Enzyme Digestion
 
-
*p-2-(1)(4)  E.X
 
-
**PCR purification kit
 
-
*p-2-(1)(7)(8)(10)  E.S
 
-
**Gel extraction
 
-
*4-(1)(2)(3)
 
-
**Colony PCR
 
-
**Insert check
 
 +
===Tuesday, 29 September===
 +
====To Do====
 +
*sig-GFP generator
 +
**PCR(the second PCR)
 +
*sig-EGFP generator(HeLa)
 +
**phosphorylation signal sequence primer
 +
**annealing signal sequence primer
 +
**Restriction Enzyme Digestion(EcoR1 + Xho1)
 +
**ligation
 +
**transformation
 +
====Results====
 +
No colony was observed (090930)
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{{:Team:Kyoto/Pad_between_dates}}
-
===Wednesday, 16 September===
+
 
 +
===Wednesday, 30 September===
====To Do====
====To Do====
 +
*sig-GFP generator
 +
**miniprep I712074(T7 promoter)
 +
**Restriction Enzyme Digestion
 +
**ligation
 +
**transformation
 +
*sig-EGFP generator(HeLa)
**ligation
**ligation
**transformation
**transformation
====Results====
====Results====
-
=====ligation=====
 
-
{| class="table"
 
-
|+conc.
 
-
!NO.!!Sample name!!conc./(ng/ul)
 
-
|-
 
-
|1||11(stu1)||103.2
 
-
|}
 
-
 
=====Transformation=====
=====Transformation=====
-
No colony was observed (090917 10:00)　
+
Too many colonies were observed that we could find no fignificant differences between the treated cluture the control. (091001)
 +
{{:Team:Kyoto/Pad_between_dates}}
 +
===Thursday, 01 October===
 +
====To Do====
 +
No experiments were undertaken
 +
{{:Team:Kyoto/Pad_between_dates}}
 +
 +
===Friday, 02 October===
 +
====To Do====
 +
No experiments were undertaken
 +
{{:Team:Kyoto/Pad_between_dates}}
 +
 +
===Saturday, 03 October===
 +
====To Do====
 +
No experiments were undertaken
 +
{{:Team:Kyoto/Pad_between_dates}}
 +
 +
===Saturday, 04 October===
 +
====To Do====
 +
No experiments were undertaken
{{:Team:Kyoto/Pad_between_dates}}
{{:Team:Kyoto/Pad_between_dates}}
-
===Thursday, 17 September===
+
===Sunday, 05 October===
====To Do====
====To Do====
-
*Make repetitive sequence (MPR)
+
No experiments were undertaken
-
*Transformation(retry)
+
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Latest revision as of 15:00, 21 October 2009

  1. Home
  2. CiC
  3. Notebook

0907-0911
constructing HIV-TAT-(LALAAAA)3-generator
0914-0918
constructing HIV-TAT-(LALAAAA)3-generator
0921-0925
constructing sig-GFP generator,sig-EGFP generator(HeLa)
0928-1002
constructing sig-GFP generator,sig-EGFP generator(HeLa)
1005-1009
constructing sig-GFP generator,sig-EGFP generator(HeLa)
1012-1016
Observation
1019-1023
Observation

0928-1004 : constructing sig-GFP generator and sig-EGFP generator(HeLa)

Monday, 28 September

To Do

  • sig-EGFP generator(HeLa)
    • Restriction Enzyme Digestion(Hind3)

Results

They were unexpectedly digested by Hind3,the Restriction Enzyme which would not work if the signal sequence were inserted in the vector correctly.

Tuesday, 29 September

To Do

  • sig-GFP generator
    • PCR(the second PCR)
  • sig-EGFP generator(HeLa)
    • phosphorylation signal sequence primer
    • annealing signal sequence primer
    • Restriction Enzyme Digestion(EcoR1 + Xho1)
    • ligation
    • transformation

Results

No colony was observed (090930)

Wednesday, 30 September

To Do

  • sig-GFP generator
    • miniprep I712074(T7 promoter)
    • Restriction Enzyme Digestion
    • ligation
    • transformation
  • sig-EGFP generator(HeLa)
    • ligation
    • transformation

Results

Transformation

Too many colonies were observed that we could find no fignificant differences between the treated cluture the control. (091001)

Thursday, 01 October

To Do

No experiments were undertaken

Friday, 02 October

To Do

No experiments were undertaken

Saturday, 03 October

To Do

No experiments were undertaken

Saturday, 04 October

To Do

No experiments were undertaken

Sunday, 05 October

To Do

No experiments were undertaken

Retrieved from "http://2009.igem.org/Team:Kyoto/CiC/Notebook/0928-1002"