User:Wesley J. Houston/Notebook/MAP/2013/03/08: Difference between revisions

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* Attempted to confirm ligation by streaking the colonies on an agar plate.
* Attempted to confirm ligation by streaking the colonies on an agar plate.
* All attempts proved unsuccessful.
* All attempts proved unsuccessful.
{| 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 (Pst1) || 1.0
|-
| Enzyme 2 (Spe1 or Xba1) || 1.0
|-
| dH<sub>2</sub>O || 5.0 μL
|-
| &nbsp; || 30 μL --> 37°C/ ~15 min.
|}




Revision as of 22:02, 25 March 2013

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3/8/13

  • Ligation was repeated.
  • Attempted to grow more liquid cultures of ligated plasmids.
  • Attempted to confirm ligation by streaking the colonies on an agar plate.
  • All attempts proved unsuccessful.
Reagent Volume   Results of the ligation electrophoresis
15 μL/lane, 1% agarose; Ladder
DNA (plasmid) 20 μL
10x buffer 3.0
Enzyme 1 (Pst1) 1.0
Enzyme 2 (Spe1 or Xba1) 1.0
dH2O 5.0 μL
  30 μL --> 37°C/ ~15 min.



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