User:Wesley J. Houston/Notebook/MAP/2013/03/07

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(3/7/13)
Current revision (00:01, 26 March 2013) (view source)
(3/7/13)
 
Line 20: Line 20:
| 10x buffer || 3.0
| 10x buffer || 3.0
|-
|-
-
| enzyme 1 || 1.0
+
| EcoR1 || 1.0
|-
|-
-
| enzyme 2 || 1.0
+
| Pst1 || 1.0
|-
|-
| dH<sub>2</sub>O || 5.0 μL
| dH<sub>2</sub>O || 5.0 μL
Line 31: Line 31:
* No DNA present, will repeat ligation.
* No DNA present, will repeat ligation.
 +
** Determined to be a false positive.
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3/7/13

  • Prepped plasmids using Zyppy miniprep kit.
  • Digested plasmids with EcoR1 and Pst1.
  • Ran gel electrophoresis to confirm assembly.
Reagent Volume   Results of the ligation electrophoresis
15 μL/lane, 1% agarose; Ladder
DNA (plasmid) 20 μL
10x buffer 3.0
EcoR1 1.0
Pst1 1.0
dH2O 5.0 μL
  30 μL --> 37°C/ ~15 min.


  • No DNA present, will repeat ligation.
    • Determined to be a false positive.


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