Team:Kyoto/CiC/Notebook/1019-1023

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==0914-0918 : Plasmid Construction of HIV-TAT-(LALAAAA)3-generator ==
+
==1019-1025 :Obserbation  ==
 +
 
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-
===Monday, 14 August===
+
===Monday, 19 October===
====To Do====
====To Do====
-
*p-2-(1)(4)(7)(8)(10)
+
* subgoal A --- Confirmation of function of TAT-liposome
-
**Miniprep
+
**observation by microscope
-
**Restriction Enzyme Digestion
+
* Subgoal B --- Liposomes with Mitochondrial Translocases
-
*p-2-(1)(4)  E.X
+
**Mitochondoria isolation
-
**PCR purification kit
+
**expressing sig-GFP in vitro with mitochondoria
-
*p-2-(1)(7)(8)(10)  E.S
+
* Subgoal C --- sig-EGFP expression in HeLa
-
**Gel extraction
+
**transfomation
-
*4-(1)(2)(3)
+
-
**Colony PCR
+
-
**Insert check
+
====Results====
====Results====
-
=====p-2-(1)(4)(7)(8)(10)=====
+
=====subgoal A --- Confirmation of function of TAT-liposome=====
-
=====p-2-(1)(7)(8)(10)  E.S=====
+
Liposome was taken by HeLa cells.See the [[Team:Kyoto/CiC/Results|results]].  
-
*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
+
-
|}
+
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=====4-(1)(2)(3)=====
+
=====Subgoal C --- sig-EGFP expression in HeLa=====
-
[[Image:Kyoto_0914_PCR_2.png|250px]]
+
sig-EGFP　was made a localization in mitochondoria.
-
*This result indicates that (1)a, (2)b and (3)a were inserted properly.
+
See the [[Team:Kyoto/CiC/Results|results]].  
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===Tuesday, 15 September===
 
-
====To Do====
 
-
'''Further to 14 Sept.'''
 
-
*p-2-(1)(4)(7)(8)(10)
+
===Tuesday, 20 October===
-
**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_start_dates}}
+
-
===Monday, 14 August===
+
====To Do====
====To Do====
-
*p-2-(1)(4)(7)(8)(10)
+
* Subgoal B --- Liposomes with Mitochondrial Translocases
-
**Miniprep
+
**observation with microscope
-
**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)=====
+
See the [[Team:Kyoto/CiC/Results|results]].  
-
=====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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{{:Team:Kyoto/Pad_between_dates}}
-
{{:Team:Kyoto/Pad_between_dates}}
+
===Wednesday, 21 October===
-
===Wednesday, 16 September===
+
====To Do====
====To Do====
-
**ligation
+
* subgoal A --- Confirmation of function of TAT-liposome
-
**transformation
+
**making liposome by natural swelling method and extruder
-
 
+
**add liposome to HeLa cells
====Results====
====Results====
-
=====ligation=====
+
According to the expectation,HIV-TAT-(LALAAAA)3-generator may work.
-
{| class="table"
+
Liposome is taken by HeLa Cells.
-
|+conc.
+
See the [[Team:Kyoto/CiC/Results|results]].  
-
!NO.!!Sample name!!conc./(ng/ul)
+
-
|-
+
-
|1||11(stu1)||103.2
+
-
|}
+
-
=====Transformation=====
 
-
No colony was observed (090917 10:00)　
 
-
 
-
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-
 
-
===Thursday, 17 September===
 
-
====To Do====
 
-
*Make repetitive sequence (MPR)
 
-
*Transformation(retry)
 
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Latest revision as of 03:08, 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

1019-1025 :Obserbation

Monday, 19 October

To Do

  • subgoal A --- Confirmation of function of TAT-liposome
    • observation by microscope
  • Subgoal B --- Liposomes with Mitochondrial Translocases
    • Mitochondoria isolation
    • expressing sig-GFP in vitro with mitochondoria
  • Subgoal C --- sig-EGFP expression in HeLa
    • transfomation

Results

subgoal A --- Confirmation of function of TAT-liposome

Liposome was taken by HeLa cells.See the results.

Subgoal C --- sig-EGFP expression in HeLa

sig-EGFP　was made a localization in mitochondoria. See the results.

Tuesday, 20 October

To Do

  • Subgoal B --- Liposomes with Mitochondrial Translocases
    • observation with microscope

Results

See the results.

Wednesday, 21 October

To Do

  • subgoal A --- Confirmation of function of TAT-liposome
    • making liposome by natural swelling method and extruder
    • add liposome to HeLa cells

Results

According to the expectation,HIV-TAT-(LALAAAA)3-generator may work. Liposome is taken by HeLa Cells. See the results.

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