Difference between revisions of "User:Wesley J. Houston/Notebook/MAP/2013/03/22"

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==3/22/13==
 
==3/22/13==
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* Gel purification was performed using the KAH201 plasmids prepped on 2/28, as well as the KAH182 plasmids prepped on 3/21.
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** No DNA was present in the second lane, and the concentrations and purities were determined to be too low to complete a ligation.
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* Grew more KAH201 liquid cultures.
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** Placed colonies in LB broth overnight in shaking 37C incubator.
  
 
{| class="wikitable" border="0" cellspacing="3" <!-- Digest rxn. table -->
 
{| class="wikitable" border="0" cellspacing="3" <!-- Digest rxn. table -->
 
|-valign="top"
 
|-valign="top"
 
| <u>Reagent</u> || <u>Volume</u> || &nbsp;
 
| <u>Reagent</u> || <u>Volume</u> || &nbsp;
| rowspan="9" | [[Image:Electrophoresis4.jpg‎ |150px|Results of third electrophoresis]]<br>15 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
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| rowspan="9" | [[Image:Electrophoresis4.jpg‎ |150px|Results of third electrophoresis]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
 
|-
 
|-
 
| DNA (plasmid) || 20 μL
 
| DNA (plasmid) || 20 μL
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| &nbsp; || 30 μL --> 37°C/ ~15 min.
 
| &nbsp; || 30 μL --> 37°C/ ~15 min.
 
|}
 
|}
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> Measure conc.'s of gel purified fragments
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{| class="wikitable" border="0" cellspacing="3" <!-- [DNA] table -->
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|-
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| <u>Sample</u> || <u>260/280</u> || <u>ng/μL</u>
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|-
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| 1. KAH201 (S/P) || 0.32 || 0.851
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|-
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| 2. KAH182 (X/P) || 0.385 || 1.064
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|}
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Revision as of 22:23, 25 March 2013

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3/22/13

  • Gel purification was performed using the KAH201 plasmids prepped on 2/28, as well as the KAH182 plasmids prepped on 3/21.
    • No DNA was present in the second lane, and the concentrations and purities were determined to be too low to complete a ligation.
  • Grew more KAH201 liquid cultures.
    • Placed colonies in LB broth overnight in shaking 37C incubator.
Reagent Volume   Results of third electrophoresis
30 μL/lane, 1% agarose; Ladder
DNA (plasmid) 20 μL
10x buffer 3.0
enzyme 1 (Pst1) 1.0
enzyme 2 (Xba1 or Spe1) 1.0
dH2O 5.0 μL
  30 μL --> 37°C/ ~15 min.

> Measure conc.'s of gel purified fragments

Sample 260/280 ng/μL
1. KAH201 (S/P) 0.32 0.851
2. KAH182 (X/P) 0.385 1.064