User:Karmella Haynes/Notebook/BioBrick cloning/2013/04/01

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(04/01/13)
 
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| bgcolor=#cfcfcf | Reagent
| bgcolor=#cfcfcf | Reagent
| bgcolor=#cfcfcf | Volume  
| bgcolor=#cfcfcf | Volume  
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| rowspan="7" | <u>Expected:</u><br>1. BB 1 = size<br>2. BB2 = size<br>
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| rowspan="7" | <u>Expected:</u><br>1-4. MV9 = 4531, 80
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| rowspan="7" | [[Image:KAH040113.jpg|200px|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
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| rowspan="7" | [[Image:KAH040113_gel1.jpg|200px|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
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| DNA(plasmid) || 3.0 μL
| DNA(plasmid) || 3.0 μL
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* Result is unclear for #1. Submit for sequencing.
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Current revision

Karmella's BioBrick Cloning Main project page
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04/01/13

  • Check MV9 clones



MV9 clones

  • Picked colonies from last cloning attempt. Streaked on LB amp agar.
  • Grew up 2 mL cultures overnight and did minipreps.


Minipreps

  • Check with S/X digests
    • S present in upstream cloning sites, X inside the new insert
Reagent Volume Expected:
1-4. MV9 = 4531, 80
Hover name
15 μL/lane; 1% agarose; Ladder
DNA(plasmid) 3.0 μL
10X buffer 1.5
EcoRI 1.0
PstI 1.0
dH2O 8.5
  15 μL --> 37°C/ ~15 min.


  • Result is unclear for #1. Submit for sequencing.


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