Team:Kyoto/CiC/Notebook/0907-0911

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==0914-0918 : Plasmid Construction of HIV-TAT-(LALAAAA)3-generator ==
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==097-0911 : Plasmid Construction of HIV-TAT-(LALAAAA)3-generator ==
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===Monday, 14 August===
+
===Wednesday, 16 September===
====To Do====
====To Do====
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*p-2-(1)(4)(7)(8)(10)
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*HIV-TAT-(LALAAAA)3
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**Miniprep
+
**PCR
-
**Restriction Enzyme Digestion
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**Restriction Enzyme Digestion(EcoR1 + Spe1)
-
*p-2-(1)(4)  E.X
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*B0015
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**PCR purification kit
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**Restriction Enzyme Digestion(EcoR1 + Xba1)
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*p-2-(1)(7)(8)(10)  E.S
+
-
**Gel extraction
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*4-(1)(2)(3)
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**Colony PCR
+
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**Insert check
+
====Results====
====Results====
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=====p-2-(1)(4)(7)(8)(10)=====
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*Band of expected size were observed.
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=====p-2-(1)(7)(8)(10)  E.S=====
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*Gel extraction kit
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[[Image:Kyoto_0914_PCR.png|250px]]
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{| class="table"
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|+Miniprep product conc.
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!NO.!!Sample name!!conc./(ng/ul)
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-
|-
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|1||p-2-(1)||9.3
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-
|-
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|2||p-2-(7)||25.9
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|-
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|3||p-2-(8)||8.6
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-
|-
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|4||p-2-(10)||7.5
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-
|}
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=====4-(1)(2)(3)=====
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[[Image:Kyoto_0914_PCR_2.png|250px]]
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*This result indicates that (1)a, (2)b and (3)a were inserted properly.
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===Tuesday, 15 September===
 
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====To Do====
 
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'''Further to 14 Sept.'''
 
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*p-2-(1)(4)(7)(8)(10)
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===Fryday, 11 September===
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**Miniprep
+
-
**Restriction Enzyme Digestion
+
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*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
+
-
 
+
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{{:Team:Kyoto/Pad_between_dates}}
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===Wednesday, 16 September===
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====To Do====
====To Do====
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**ligation
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*HIV-TAT-(LALAAAA)3
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**transformation
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**Ligation
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**Transformation
====Results====
====Results====
=====ligation=====
=====ligation=====
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!NO.!!Sample name!!conc./(ng/ul)
!NO.!!Sample name!!conc./(ng/ul)
|-
|-
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|1||11(stu1)||103.2
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|1||HIV-TAT sample1(EcoR1,Spe1)||36.2
 +
|-
 +
|2||HIV-TAT sample2(EcoR1,Spe1)||21.7
 +
|-
 +
|3||HIV-TAT sample3(EcoR1,Spe1)||47.9
 +
|-
 +
|4||B0015(EcoR1,Xba1)||2.48
|}
|}
=====Transformation=====
=====Transformation=====
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No colony was observed (090917 10:00)　
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Colony was observed (090912)　
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{{:Team:Kyoto/Pad_between_dates}}
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===Thursday, 17 September===
 
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====To Do====
 
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*Make repetitive sequence (MPR)
 
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*Transformation(retry)
 
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{{:Team:Kyoto/Pad_endof_dates}}
 
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<html></div><!-- /#rightcontents --></html>
 

Latest revision as of 16:15, 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

097-0911 : Plasmid Construction of HIV-TAT-(LALAAAA)3-generator

Wednesday, 16 September

To Do

  • HIV-TAT-(LALAAAA)3
    • PCR
    • Restriction Enzyme Digestion(EcoR1 + Spe1)
  • B0015
    • Restriction Enzyme Digestion(EcoR1 + Xba1)

Results

  • Band of expected size were observed.

Fryday, 11 September

To Do

  • HIV-TAT-(LALAAAA)3
    • Ligation
    • Transformation

Results

ligation
conc.
NO.Sample nameconc./(ng/ul)
1HIV-TAT sample1(EcoR1,Spe1)36.2
2HIV-TAT sample2(EcoR1,Spe1)21.7
3HIV-TAT sample3(EcoR1,Spe1)47.9
4B0015(EcoR1,Xba1)2.48
Transformation

Colony was observed (090912)　

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