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

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* Used bit of streak for PCR today
* Used bit of streak for PCR today
* All parts are on pSB1A3-based vectors (http://partsregistry.org/Part:pSB1A3)
* All parts are on pSB1A3-based vectors (http://partsregistry.org/Part:pSB1A3)
 +
** Use primers VF2 and VR to amplify inserts
{| {{table}} border="1" cellspacing="3" <!-- Digest check rxn. table -->
{| {{table}} border="1" cellspacing="3" <!-- Digest check rxn. table -->

Revision as of 21:01, 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
  • All parts are on pSB1A3-based vectors (http://partsregistry.org/Part:pSB1A3)
    • Use primers VF2 and VR to amplify inserts
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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