Difference between revisions of "User:Karmella Haynes/Notebook/BioBrick cloning/2013/01/11"

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----
 
----
'''Minipreps'''<br>
+
'''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
| 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]
+
| 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]
 
|-
 
|-
 
| DNA(plasmid) || 3.0 μL
 
| DNA(plasmid) || 3.0 μL
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|-
 
|-
 
| dH<sub>2</sub>O || 8.5
 
| dH<sub>2</sub>O || 8.5
|-
+
|- valign="top"
| &nbsp; || 15 μL --> 37°C/ ~15 min.
+
| &nbsp; || 15 μL
 
|}
 
|}
 +
*  --> 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 15:16, 11 January 2013

Owwnotebook icon.pngKarmella's BioBrick Cloning <html><img src="/images/9/94/Report.png" border="0" /></html> 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