Team:Kyoto/CiC/Notebook/0921-0925

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-
==0914-0918 : what to do. what to do. what to do. what to do. ==
+
==0914-0918 : constructing sig-GFP generator,sig-EGFP generator(HeLa)  ==
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{{:Team:Kyoto/Pad_start_dates}}
-
===Monday, 14 August===
+
===Monday, 21 September===
====To Do====
====To Do====
-
*p-2-(1)(4)(7)(8)(10)
+
*HIV-TAT-(LALAAAA)3-generator
-
**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
**Colony PCR
-
**Insert check
 
-
 
====Results====
====Results====
-
=====p-2-(1)(4)(7)(8)(10)=====
+
*Binds of expected size were observed.
-
=====p-2-(1)(7)(8)(10)  E.S=====
+
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-
*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)=====
+
===Tuesday, 22 September===
-
[[Image:Kyoto_0914_PCR_2.png|250px]]
+
====To Do====
-
*This result indicates that (1)a, (2)b and (3)a were inserted properly.
+
*HIV-TAT-(LALAAAA)3-generator
 +
**miniprep
 +
*sig-EGFP generator(HeLa)
 +
**Restriction Enzyme PEGFP-N3 VECTOR (EGFP generator)
 +
**hosphorylation signal sequence primer
 +
**annering signal sequence primer
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===Tuesday, 15 September===
 
-
====To Do====
 
-
'''Further to 14 Sept.'''
 
-
*p-2-(1)(4)(7)(8)(10)
+
===Wednesday, 23 September===
-
**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
+
-
 
+
-
{{:Team:Kyoto/Pad_between_dates}}
+
-
===Wednesday, 16 September===
+
====To Do====
====To Do====
 +
*sig-EGFP generator
**ligation
**ligation
**transformation
**transformation
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{| class="table"
{| class="table"
|+conc.
|+conc.
-
!NO.!!Sample name!!conc./(ng/ul)
+
!NO.!!Sample name!!conc./(ng/ul)!!conc.(ul)
 +
|-
 +
|1||pEGFP-N3(EcoR1,Xho1)||224.3||1
|-
|-
-
|1||11(stu1)||103.2
+
|2||signal sequence||0.3||11
|}
|}
=====Transformation=====
=====Transformation=====
-
No colony was observed (090917 10:00)　
+
No colony was observed. (090917 10:00)　
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Latest revision as of 02:06, 22 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

0914-0918 : constructing sig-GFP generator,sig-EGFP generator(HeLa)

Monday, 21 September

To Do

  • HIV-TAT-(LALAAAA)3-generator
    • Colony PCR

Results

  • Binds of expected size were observed.

Tuesday, 22 September

To Do

  • HIV-TAT-(LALAAAA)3-generator
    • miniprep
  • sig-EGFP generator(HeLa)
    • Restriction Enzyme PEGFP-N3 VECTOR (EGFP generator)
    • hosphorylation signal sequence primer
    • annering signal sequence primer

Wednesday, 23 September

To Do

  • sig-EGFP generator
    • ligation
    • transformation

Results

ligation
conc.
NO.Sample nameconc./(ng/ul)conc.(ul)
1pEGFP-N3(EcoR1,Xho1)224.31
2signal sequence0.311
Transformation

No colony was observed. (090917 10:00)　

Thursday, 17 September

To Do

  • Make repetitive sequence (MPR)
  • Transformation(retry)

Retrieved from "http://2009.igem.org/Team:Kyoto/CiC/Notebook/0921-0925"