User:Karmella Haynes/Notebook/BioBrick cloning/2012/12/03

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==mm/dd/yy==
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==12/03/12==
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* Assembly: KAH111 + C-His expression vector (trouble shooting for Behzad)
* Assembly: KAH111 + C-His expression vector (trouble shooting for Behzad)
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{| {{table}} cellspacing="3" <!-- Ligations table -->
{| {{table}} cellspacing="3" <!-- Ligations table -->
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|- bgcolor=#cfcfcf
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| Ligation || <font color="blue"><u>Plate results (lig : neg crtl)</u> mm/dd/yy</font>
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| Ligation || <font color="blue"><u>Plate results (lig : neg crtl)</u> 12/04/12</font>
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|-
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| 1. KAH111(E/P)/1123, 16 ng + C-His(E/P)/3600, 25 ng || <font color="blue">new BioBrick #:1 (Pick #)</font>
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| 1. KAH111(E/P)/1123, 16 ng + C-His(E/P)/3600, 25 ng || <font color="red">KAH111/C-His 0 colonies</font>
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|-
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| 2. C-His(E/P)/ 25 ng || &nbsp;
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| 2. C-His(E/P)/ 25 ng || 0 colonies
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{| {{table}} cellspacing="3" <!-- Ligation rxn table -->
{| {{table}} cellspacing="3" <!-- Ligation rxn table -->
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| &nbsp;            || 1    || 2    ||
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| &nbsp;            || 1    || 2     
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|-
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| Insert DNA        || 1.0  || ---  ||
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| Insert DNA        || 1.0  || ---   
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| Vector DNA        || 0.5  || 0.5  ||
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| Vector DNA        || 0.5  || 0.5   
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| 2x lgn buf (Roche) || 5.0  || 5.0  ||
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| 2x lgn buf (Roche) || 5.0  || 5.0   
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|-
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| T4 ligase (NEB)    || 1.0  || 1.0  ||
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| T4 ligase (NEB)    || 1.0  || 1.0   
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|-
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| dH<sub>2</sub>O    || 2.5  || 3.5  ||
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| dH<sub>2</sub>O    || 2.5  || 3.5   
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|-
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| &nbsp;            || 10 μL || 10 μL ||
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| &nbsp;            || 10 μL || 10 μL  
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Current revision

Karmella's BioBrick Cloning Main project page
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12/03/12

  • Assembly: KAH111 + C-His expression vector (trouble shooting for Behzad)


Assemblies

  1. KAH111/C-His: KAH111/(E/P)/1123 + C-His/(E/P)/3600


  • Digests (Fermentas FD)
    • Already done by Behzad


  • Measure conc.'s
    • Behzad: KAH111 = 15 ng/μL
    • Behzad: C-His = 57 ng/μL


  • Ligations
Ligation Plate results (lig : neg crtl) 12/04/12
1. KAH111(E/P)/1123, 16 ng + C-His(E/P)/3600, 25 ng KAH111/C-His 0 colonies
2. C-His(E/P)/ 25 ng 0 colonies
  1 2
Insert DNA 1.0 ---
Vector DNA 0.5 0.5
2x lgn buf (Roche) 5.0 5.0
T4 ligase (NEB) 1.0 1.0
dH2O 2.5 3.5
  10 μL 10 μL



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