User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2012/12/14

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(14/12/2012)
Current revision (17:51, 12 January 2013) (view source)
(14/12/2012)
 
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| Ligation || <font color="blue"><u>Plate results (lig : neg crtl)</u> 12/12/12</font>
| Ligation || <font color="blue"><u>Plate results (lig : neg crtl)</u> 12/12/12</font>
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| 1. E. coli NEB10B KAH111(E/P)/size, 16 ng + C-His(E/P)/3600, 25ng || <font color="blue">KAH111/C-His 3:1 (Pick 4)</font>  
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| 1. E. coli NEB10B KAH111(E/P)/size, 16 ng + C-His(E/P)/3600, 25ng || <font color="blue">KAH111/C-His 3:1 No Colonies</font>  
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| 2. E. coli BL-21  KAH111(E/P)/size, 16 ng + C-His(E/P)/3600, 25ng || <font color="blue">KAH111/C-His 3:1 (Pick 4)</font>  
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| 2. E. coli BL-21  KAH111(E/P)/size, 16 ng + C-His(E/P)/3600, 25ng || <font color="blue">KAH111/C-His 3:1 No Colonies</font>  
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| 3. C-His(E/P)/ 25 ng ||  
| 3. C-His(E/P)/ 25 ng ||  

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14/12/2012

Ligation of KAH111 and C-His vector in NEB10B and BL21

  • Ligations
Ligation Plate results (lig : neg crtl) 12/12/12
1. E. coli NEB10B KAH111(E/P)/size, 16 ng + C-His(E/P)/3600, 25ng KAH111/C-His 3:1 No Colonies
2. E. coli BL-21 KAH111(E/P)/size, 16 ng + C-His(E/P)/3600, 25ng KAH111/C-His 3:1 No Colonies
3. C-His(E/P)/ 25 ng
  1 2 3
Insert DNA 3.12 3.12 ---
Vector DNA 2.94 2.94 2.94
2x lgn buf (Roche) 7.06 7.06 5.0
T4 ligase (NEB) 1.0 1.0 1.0
dH2O 0 0 0.88
  14.12 μL 14.12 μL 10μl
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