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

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Current revision (00:01, 26 March 2013) (view source)
(3/7/13)
 
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==3/7/13==
==3/7/13==
* Prepped plasmids using Zyppy miniprep kit.
* Prepped plasmids using Zyppy miniprep kit.
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* Digested plasmids with EcoR1 and Pst1 (table will be made).
+
* Digested plasmids with EcoR1 and Pst1.
-
* Ran gel electrophoresis to confirm assembly (picture will be uploaded).
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* Ran gel electrophoresis to confirm assembly.
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{| class="wikitable" border="0" cellspacing="3" <!-- Digest rxn. table -->
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|-valign="top"
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| <u>Reagent</u> || <u>Volume</u> || &nbsp;
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| rowspan="9" | [[Image:Electrophoresis3.jpg‎ |150px|Results of the ligation electrophoresis]]<br>15 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
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|-
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| DNA (plasmid) || 20 μL
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|-
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| 10x buffer || 3.0
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|-
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| EcoR1 || 1.0
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|-
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| Pst1 || 1.0
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|-
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| dH<sub>2</sub>O || 5.0 μL
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|-
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| &nbsp; || 30 μL --> 37°C/ ~15 min.
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|}
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* 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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