Difference between revisions of "Haynes Lab:Notebook/David Barclay Undergrad Training/Plasmid Transformation/2013/10/03"

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(Continuation of Restriction Digests and Gel Electrophoresis)
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Haynes BioBrick Cloning</span>
 
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Haynes BioBrick Cloning</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>}}
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|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 | Volume
 
| bgcolor=#cfcfcf | Volume
 
| rowspan="7" | <u>Expected:</u><br>1. fshPCD = 186 bp, 4968 bp<br>
 
| rowspan="7" | <u>Expected:</u><br>1. fshPCD = 186 bp, 4968 bp<br>
| rowspan="7" | [[Image:IMAG0329.jpg|400px|Today's gel]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
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| rowspan="7" | [[Image:Gel DKB 10-3-13.jpg|400px|Today's gel]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
 
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| DNA(plasmid) || 5.0 μL
 
| DNA(plasmid) || 5.0 μL
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'''Conclusion'''
 
'''Conclusion'''
 
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There were faint bands at the 186 position; however, they are harder to see on the picture. The 4968 bands for all 3 are very visible. Fairly successful digest.
  
 
'''DNA Sequencing Samples'''
 
'''DNA Sequencing Samples'''
 
<!-- * signs create an automatically bulleted list. Replace 'date' with the date you submitted the samples -->
 
<!-- * signs create an automatically bulleted list. Replace 'date' with the date you submitted the samples -->
 
<!-- The Order Number is shown at the top of your sequencing order form. Record in the indicated spot below -->
 
<!-- The Order Number is shown at the top of your sequencing order form. Record in the indicated spot below -->
* Submitted to DNASU on:  
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* Submitted to DNASU on: 10/7/2013
* Order Number:  
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* Order Number: 8213
 
<!-- # signs create an automatically numbered list. Replace Plasmid 1 & 2 with the names of your plasmids. Replace 'name' with the appropriate name of each primer. -->
 
<!-- # signs create an automatically numbered list. Replace Plasmid 1 & 2 with the names of your plasmids. Replace 'name' with the appropriate name of each primer. -->
 
# Plasmid 1 (fshPCD) - Forward P001
 
# Plasmid 1 (fshPCD) - Forward P001

Latest revision as of 22:24, 26 September 2017

Owwnotebook icon.png Haynes BioBrick Cloning Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

Continuation of Restriction Digests and Gel Electrophoresis

Restriction Digest Table

  • Checked plasmid minipreps with EcoRI/PstI digests
Reagent Volume Expected:
1. fshPCD = 186 bp, 4968 bp
Today's gel
15 μL/lane; 1% agarose; Ladder
DNA(plasmid) 5.0 μL
10X FD Buffer 1.5
EcoRI 1.0
PstI 1.0
dH2O 6.5
  15 μL --> 37°C/ 15 min.

Conclusion There were faint bands at the 186 position; however, they are harder to see on the picture. The 4968 bands for all 3 are very visible. Fairly successful digest.

DNA Sequencing Samples

  • Submitted to DNASU on: 10/7/2013
  • Order Number: 8213
  1. Plasmid 1 (fshPCD) - Forward P001
  2. Plasmid 1 (fshPCD) - Reverse P002