User:Karmella Haynes/Notebook/BioBrick cloning/2013/01/11

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(01/11/13)
(01/11/13)
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| dH<sub>2</sub>O || 8.5
| dH<sub>2</sub>O || 8.5
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| &nbsp; || 15 μL --> 37°C/ ~15 min.
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| &nbsp; || 15 μL
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*  --> 37°C/ ~15 min.
* None of the digests show both expected bands; probably just vector
* None of the digests show both expected bands; probably just vector

Revision as of 17:07, 11 January 2013

Karmella's BioBrick Cloning Main project page
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01/11/13

  • Golden Gate assembly trial 1: mini preps/ digests
  • Golden gate assembly trial 2: transformation



Minipreps

  • Check with E/P digests
Reagent Volume Expected:
1-4. hPCD-BL01/pSB1A3 = 2686, 2000
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.
  • None of the digests show both expected bands; probably just vector



Golden gate assembly trial 2

  • Add 10.0 μL assembly reaction to 50.0 μL DH5α-Turbo; ice/ 5 min.; plate on amp agar



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