User:Karmella Haynes/Notebook/BioBrick cloning/2013/03/06

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| bgcolor=#cfcfcf | Volume  
| bgcolor=#cfcfcf | Volume  
| rowspan="7" | <u>Expected:</u><br>Primers add ~250bp<br>1-3. BB 1 (# bp) = ##<br>4-6. R007.. (# bp) = ##<br>7. K.. (# bp) = ##
| rowspan="7" | <u>Expected:</u><br>Primers add ~250bp<br>1-3. BB 1 (# bp) = ##<br>4-6. R007.. (# bp) = ##<br>7. K.. (# bp) = ##
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| rowspan="7" | [[Image:KAH030713_gel1.jpg|250px|PCR gel]]<br>10 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
+
| rowspan="7" | [[Image:KAH030713_gel1.jpg|300px|PCR gel]]<br>10 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
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| Bacteria from tip || ---
| Bacteria from tip || ---

Revision as of 20:58, 7 March 2013

Karmella's BioBrick Cloning Main project page
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03/06/13

  • Colony PCR: for Rene's project


Colony PCR

  • Streaked single colonies on gridded plates yesterday
  • Used bit of streak for PCR today
Reagent Volume Expected:
Primers add ~250bp
1-3. BB 1 (# bp) = ##
4-6. R007.. (# bp) = ##
7. K.. (# bp) = ## 		PCR gel
10 μL/lane; 1% agarose; Ladder
Bacteria from tip ---
10 μM VF2 primer 1.0
10 μM VR primer 1.0
2x GoTaq Green 12.5
dH2O 10.5
  25.0

PCR

  • 95°C, 3 min
  • [95°C, 15 sec; 57°C 15 sec; 72 °C 60 sec] x35
  • 72°C, 3 min
  • 4°C, ∞


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