User:Jose Fabricio Lopez/Notebook/Logbook/2011/08/23: Difference between revisions

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=Objectives=
=Objectives=
* Gel extraccion of [[User:Jose Fabricio Lopez/Notebook/Logbook/2011/08/19]]
* Gel extraccion of [[User:Jose Fabricio Lopez/Notebook/Logbook/2011/08/19| PCRs]]
* PCR for Insert. Again! Low concentration in gel extraction.


==Gel extraction==
==Gel extraction==
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I need to repeat the insert PCR and make a new Gel extraction for that biopart.
I need to repeat the insert PCR and make a new Gel extraction for that biopart.
==PCR for inserts==
{| border=1
!Reagents
!PCR 1 [http://partsregistry.org/wiki/index.php?title=Part:BBa_J04450 P1A7/2011]
!PCR 2 [http://partsregistry.org/wiki/index.php?title=Part:BBa_J04450 P1A7/2011]
!PCR 3 [http://partsregistry.org/wiki/index.php?title=Part:BBa_J04450 P1A7/2011](Control+)
|--
|DNA
|1μL
|1μL
|1μL
|--
|Buffer 5x PhusionHF
|10μL
|10μL
|10μL
|--
|dNTPs(8mM)
|5μL
|5μL
|5μL
|--
|Oligo(Extnd_Pref_Fwd)
|5μL
|5μL
|5μL**
|--
|Oligo(Extnd_Suff_Rev)
|5μL
|5μL
|5μL**
|--
|Phusion
|.5μL
|.5μL
|.5μL
|--
|H<sub>2</sub>O
|23.5μL
|23.5μL
|23.5μL
|}
**Prefix_Fwd and Suffix_Reverse Primers.


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Revision as of 16:58, 24 August 2011

iGEM 2011 <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
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Objectives

  • Gel extraccion of PCRs
  • PCR for Insert. Again! Low concentration in gel extraction.

Gel extraction

  • Quiagen Gel Extraction Kit, with elution 30μL.
  • Lane 1 Ladder.
  • Lane 2 Insert with Prefix and suffix Standard for Gibson
  • Lane 3 Backbone with Prefix and suffix complete.

I need to repeat the insert PCR and make a new Gel extraction for that biopart.


PCR for inserts

Reagents PCR 1 P1A7/2011 PCR 2 P1A7/2011 PCR 3 P1A7/2011(Control+)
DNA 1μL 1μL 1μL
Buffer 5x PhusionHF 10μL 10μL 10μL
dNTPs(8mM) 5μL 5μL 5μL
Oligo(Extnd_Pref_Fwd) 5μL 5μL 5μL**
Oligo(Extnd_Suff_Rev) 5μL 5μL 5μL**
Phusion .5μL .5μL .5μL
H2O 23.5μL 23.5μL 23.5μL
    • Prefix_Fwd and Suffix_Reverse Primers.
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