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

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'''Minipreps'''<br>
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'''Golden gate trial 1, Minipreps'''<br>
* Check with E/P digests
* Check with E/P digests
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| bgcolor=#cfcfcf | Volume  
| bgcolor=#cfcfcf | Volume  
| rowspan="7" | <u>Expected:</u><br>1-4. hPCD-BL01/pSB1A3 = 2686, 2000
| rowspan="7" | <u>Expected:</u><br>1-4. hPCD-BL01/pSB1A3 = 2686, 2000
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| rowspan="7" |  [[Image:KAH011113_gel1.jpg‎|300px|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:KAH011113_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

Current revision

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


Golden gate trial 1, 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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