Team:Kyoto/CiC/Notebook/1012-1016

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-
==0914-0918 : what to do. what to do. what to do. what to do. ==
+
==1012-1018 : Obserbation  ==
 +
 
{{:Team:Kyoto/Pad_start_dates}}
{{:Team:Kyoto/Pad_start_dates}}
-
===Monday, 14 August===
+
===Monday, 12 October===
====To Do====
====To Do====
-
*p-2-(1)(4)(7)(8)(10)
+
No experiments were undertaken
-
**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
+
-
 
+
====Results====
====Results====
-
=====p-2-(1)(4)(7)(8)(10)=====
 
-
=====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.
 
-
 
{{:Team:Kyoto/Pad_between_dates}}
{{:Team:Kyoto/Pad_between_dates}}
-
===Tuesday, 15 September===
+
 
 +
===Tuesday, 13 October===
====To Do====
====To Do====
-
'''Further to 14 Sept.'''
+
*sequence
-
 
+
**sig-GFP generator
-
*p-2-(1)(4)(7)(8)(10)
+
**sig-EGFP generator(HeLa)  
-
**Miniprep
+
====Results====
-
**Restriction Enzyme Digestion
+
*We assured that the plasmid include the correct sequence by analyzing it with sequencer.(091015)
-
*p-2-(1)(4)  E.X
+
{{:Team:Kyoto/Pad_between_dates}}
-
**PCR purification kit
+
-
*p-2-(1)(7)(8)(10)  E.S
+
-
**Gel extraction
+
-
*4-(1)(2)(3)  
+
-
**Colony PCR
+
-
**Insert check
+
 +
===Wednesday, 14 October===
 +
====To Do====
 +
No experiments were undertaken
{{:Team:Kyoto/Pad_between_dates}}
{{:Team:Kyoto/Pad_between_dates}}
-
===Wednesday, 16 September===
+
 
 +
===Thursday, 15 October===
====To Do====
====To Do====
-
**ligation
+
*Experiments to approach subgoalA
-
**transformation
+
**Making of liposome by natural swelling method and extruder
 +
{{:Team:Kyoto/Pad_between_dates}}
 +
===Friday, 16 October===
 +
====To Do====
 +
*Experiments to approach subgoalA
 +
**Making of liposome by natural swelling method and extruder
 +
**expression in vitro HIV-TAT-(LALAAAA)3-generator
 +
**add liposome to HeLa cells
====Results====
====Results====
-
=====ligation=====
+
No NBD-PE signal was observed in HeLa cells.
-
{| class="table"
+
This might be because we did not collect the right fraction after density gradient centrifugation and thus lost the liposomes.
-
|+conc.
+
{{:Team:Kyoto/Pad_between_dates}}
-
!NO.!!Sample name!!conc./(ng/ul)
+
-
|-
+
-
|1||11(stu1)||103.2
+
-
|}
+
-
=====Transformation=====
+
===Saturday, 17 October===
-
No colony was observed (090917 10:00)　
+
====To Do====
 +
*Experiments to approach subgoalA
 +
**add liposome including synthesized HIV-TAT-(LALAAAA)3 peptides to HeLa cells
 +
====Results====
 +
No NBD-PE signal was observed in HeLa cells.
{{:Team:Kyoto/Pad_between_dates}}
{{:Team:Kyoto/Pad_between_dates}}
-
===Thursday, 17 September===
+
===Sunday, 18 October===
====To Do====
====To Do====
-
*Make repetitive sequence (MPR)
+
*observation by microscope
-
*Transformation(retry)
+
====Results====
 +
We could't confirm that liposome was not taken by HeLa cells.
{{:Team:Kyoto/Pad_endof_dates}}
{{:Team:Kyoto/Pad_endof_dates}}
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Latest revision as of 20:31, 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

1012-1018 : Obserbation

Monday, 12 October

To Do

No experiments were undertaken

Results

Tuesday, 13 October

To Do

  • sequence
    • sig-GFP generator
    • sig-EGFP generator(HeLa)

Results

  • We assured that the plasmid include the correct sequence by analyzing it with sequencer.(091015)

Wednesday, 14 October

To Do

No experiments were undertaken

Thursday, 15 October

To Do

  • Experiments to approach subgoalA
    • Making of liposome by natural swelling method and extruder

Friday, 16 October

To Do

  • Experiments to approach subgoalA
    • Making of liposome by natural swelling method and extruder
    • expression in vitro HIV-TAT-(LALAAAA)3-generator
    • add liposome to HeLa cells

Results

No NBD-PE signal was observed in HeLa cells. This might be because we did not collect the right fraction after density gradient centrifugation and thus lost the liposomes.

Saturday, 17 October

To Do

  • Experiments to approach subgoalA
    • add liposome including synthesized HIV-TAT-(LALAAAA)3 peptides to HeLa cells

Results

No NBD-PE signal was observed in HeLa cells.

Sunday, 18 October

To Do

  • observation by microscope

Results

We could't confirm that liposome was not taken by HeLa cells.

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