IGEM:Slovenia HS 2015/2009/Notebook/Test/2015/07/13: Difference between revisions

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|style="background-color: #EEE"|[[Image:igem-logo-150px.png|150px]]<span style="font-size:22px;"> iGEM Project name 1</span>
|style="background-color: #EEE"|[[Image:igem-logo-150px.png|150px]]<span style="font-size:22px;"> iGEM Project name 1</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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==Plasmid isolation from medium==
==Plasmid isolation from medium==
* Isolation was conducted accrdingly to [http://openwetware.org/wiki/FCCT_Biochemistry_Lab: Plasmid DNA purification using centrifuges]<br>
* Isolation was conducted accrdingly to [http://openwetware.org/wiki/FCCT_Biochemistry_Lab: Plasmid DNA purification using centrifuges]<br>
==Absorbance of constructs==
To control how successful plasmid isolation was, we decided to measure absorbance of genes with nanodrop.
::{| border="1" cellpadding="2" cellspacing="0"
!component||||quantity (µL)||
|-
|oligo 1 (10µM)||1||10||
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|oligo 2 (10µM)||1||10||
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|dNTP (2mM)||1 ||10||
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|MgCl2||2||20||
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|buffer||2||20||
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|dH2O||11||110||
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*We have conducted electrophoresis accordingly to [http://openwetware.org/wiki/FCCT_Biochemistry_Lab:Agarose_gel_electrophoresis Agarose gel electrophoresis]<br>
*We have conducted electrophoresis accordingly to [http://openwetware.org/wiki/FCCT_Biochemistry_Lab:Agarose_gel_electrophoresis Agarose gel electrophoresis]<br>
*All restrictions were lain on the electrophoresis cell
*All restrictions were lain on the electrophoresis cell




Latest revision as of 01:03, 27 September 2017

iGEM Project name 1 Main project page
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Plasmid isolation from medium

Absorbance of constructs

To control how successful plasmid isolation was, we decided to measure absorbance of genes with nanodrop.

Control restriction

Mixture:

  • We have taken 5μL from each microcentrifuge tube
  • 0,55 ml of buffer
  • 0,2 μL of restriction enzyme Pst1

With the restriction we have linearized our plasmid.

Agarose gel electrophoresis


component quantity (µL)
oligo 1 (10µM) 1 10
oligo 2 (10µM) 1 10
dNTP (2mM) 1 10
MgCl2 2 20
buffer 2 20
dH2O 11 110


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