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

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==3/7/13==
==3/7/13==
* Prepped plasmids using Zyppy miniprep kit.
* Prepped plasmids using Zyppy miniprep kit.
* 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).
* Ran gel electrophoresis to confirm assembly.
* No DNA present, repeat ligation.
 
{| class="wikitable" border="0" cellspacing="3" <!-- Digest rxn. table -->
|-valign="top"
| <u>Reagent</u> || <u>Volume</u> || &nbsp;
| 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]
|-
| DNA (plasmid) || 20 μL
|-
| 10x buffer || 3.0
|-
| enzyme 1 || 1.0
|-
| enzyme 2 || 1.0
|-
| dH<sub>2</sub>O || 5.0 μL
|-
| &nbsp; || 30 μL --> 37°C/ ~15 min.
|}
 
 
* No DNA present, will repeat ligation.


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Revision as of 13:17, 18 March 2013

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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
enzyme 1 1.0
enzyme 2 1.0
dH2O 5.0 μL
  30 μL --> 37°C/ ~15 min.


  • No DNA present, will repeat ligation.


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