User:Wesley J. Houston/Notebook/MAP/2013/03/25: Difference between revisions

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
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|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
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==3/25/13==
==3/25/13==
* Prepped plasmids of KAH201 and determined concentration and purity.
* Prepped plasmids of KAH201 and determined concentration and purity.
* Followed by digestion and gel electrophoresis, then purification.
* However, concentrations were determined to be too low to complete a ligation.
* More liquid cultures of KAH182 and KAH201 were prepped.
** Colonies placed into LB broth overnight in shaking 37C incubator.


{| class="wikitable" border="0" cellspacing="3" <!-- [DNA] table -->
{| class="wikitable" border="0" cellspacing="3" <!-- [DNA] table -->
Line 44: Line 48:
| 1. KAH201 (S/P) || 0.727 || 2.54
| 1. KAH201 (S/P) || 0.727 || 2.54
|-
|-
| 2. KAH182 (X/P) || 0.789 || 13596
| 2. KAH182 (X/P) || 0.789 || 1.596
|}
|}


* Due to the low concentrations, I made more liquid cultures of KAH182 and KAH201.
** Followed the established protocol for making liquid cultures: pick one colony and place into LB broth overnight in shaking 37C incubator.


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Latest revision as of 22:34, 26 September 2017

Project name Main project page
Previous entry      Next entry

3/25/13

  • Prepped plasmids of KAH201 and determined concentration and purity.
  • Followed by digestion and gel electrophoresis, then purification.
  • However, concentrations were determined to be too low to complete a ligation.
  • More liquid cultures of KAH182 and KAH201 were prepped.
    • Colonies placed into LB broth overnight in shaking 37C incubator.
Sample 260/280 ng/μL
1. KAH201 (Plasmid) 1.795 54.054
2. KAH201 (Plasmid) 1.708 47.244


Reagent Volume   Results of the fourth electrophoresis attempt
30 μL/lane, 1% agarose; Ladder
DNA (plasmid) 25 μL
10x buffer 3.0
enzyme 1 (Pst1) 1.0
enzyme 2 (Xba1 or Spe1) 1.0
dH2O 0.0 μL
  30 μL --> 37°C/ ~15 min.

> Measure conc.'s of gel purified fragments

Sample 260/280 ng/μL
1. KAH201 (S/P) 0.727 2.54
2. KAH182 (X/P) 0.789 1.596
  • Due to the low concentrations, I made more liquid cultures of KAH182 and KAH201.
    • Followed the established protocol for making liquid cultures: pick one colony and place into LB broth overnight in shaking 37C incubator.


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