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

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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||
|-
|oligo 2 (10µM)||1||10||
|-
|dNTP (2mM)||1 ||10||
|-
|MgCl2||2||20||
|-
|buffer||2||20||
|-
|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




Revision as of 08:43, 27 July 2015

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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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