Haynes Lab:Notebook/Jan/2015/03/28: Difference between revisions

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
|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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| bgcolor=#cfcfcf | Reagent
| bgcolor=#cfcfcf | Reagent
| bgcolor=#cfcfcf | Volume
| bgcolor=#cfcfcf | Volume
| rowspan="7" | Expected:<br>* BD006/MV2 = 7207, 1243, 288<br>* MV2 only = 2998, 1243, 288
| rowspan="7" | Expected:<br>* BD006/MV2 = 7207, 1243, 288<br>* MV2 only = 2998, 1243, 288<br>* BD006/V0120 = 7036, 497
| rowspan="7" | [[Image:KAH032715_gel1.jpg|270px|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
| rowspan="7" | [[Image:KAH032815_gel1.jpg|270px|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
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| DNA (plasmid) || 3.0 μL
| DNA (plasmid) || 3.0 μL

Latest revision as of 00:52, 27 September 2017

Project name Main project page
Previous entry      Next entry

03/28/15

  • Minipreps - BD006/MV2 colonies (from 3/26/15 ligations), 5 mL each (4 total)


Minipreps - BD006/MV2 colonies

  • Sigma GenElute kit.
    • Elute & back-elute w/ 70 μL elution sln.
  1. BD006/MV2 plate 1, a
  2. BD006/MV2 plate 1, b
  3. BD006/MV2 plate 2, a
  4. BD006/MV2 plate 2, b


  • Restriction Digest check (Fermentas FD)
    • Cut with ApaLI (E/S insertion got rid of BB RFC23 cut sites)


Reagent Volume Expected:
* BD006/MV2 = 7207, 1243, 288
* MV2 only = 2998, 1243, 288
* BD006/V0120 = 7036, 497 		Hover name
30 μL/lane, 1% agarose; Ladder
DNA (plasmid) 3.0 μL
10x buffer 1.5
ApaLI 1.0
dH2O 9.5
  15 μL --> 37°C/ ~10 min.


  • Conclusions
    • Success! All four look like BD006/MV2
    • Submit for sequencing next week


  • Miniprep conc.'s
Sample OD260 260/280 ng/μL
1. BD006/MV2 1-a 0.274 1.44 274.10
2. BD006/MV2 1-b 0.299 1.93 298.68
3. BD006/MV2 2-a 0.331 1.93 330.69
4. BD006/MV2 2-b 0.306 1.90 305.94



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