User:Jose Fabricio Lopez/Notebook/Logbook/2011/06/20

From OpenWetWare

< User:Jose Fabricio Lopez | Notebook | Logbook | 2011 | 06
Revision as of 14:09, 28 June 2011 by Jose Fabricio Lopez (Talk | contribs)
(diff) ←Older revision | Current revision (diff) | Newer revision→ (diff)
Jump to: navigation, search
iGEM 2011 Main project page
Previous entry      Next entry

Objectives

  • PCR from extractions
  • Primers (5 pM/μL) (new set because we are improving new protocol of standardization).
    • Primer Forward= CGG GCC CCA TTA TTA TCA TGA CAT TAA CCT ATA AAA ATA GG
      • It includes GGGCCC. Site for ApaI.
    • Primer Reverse= CGA GCT CGC CTT TTG CTC ACA TCT TCT TTC C
      • It includes GAGCTC. Site SacI.

PCR

  • DNA was obtained from extractions and diluted(2μL DNA in 98μL H2O)

PCR amplification

Reactives Pablo´s extraction Tube 1 Tube 2
Buffer Standard Taq 10x NEB 5μL 5μL 5μL
Primer F 5μL 5μL 5μL
Primer R 5μL 5μL 5μL
10mM dNTPs 1μL 1μL 1μL
DNA 2μL 2μL 2μL
Taq .5μL .5μL .5μL
H2O 31.5 31.5 31.5
Total 50μL 50μL 50μL

Control PCR

Reactives -R -F -O
Buffer Standard Taq 10x NEB 2μL 2μL 2μL
Primer F 2μL 0μL 0μL
Primer R 0μL 2μL 0μL
.4mM dNTPs 10μL 10μL 10μL
DNA(Pablo's extraction) 2μL 2μL 2μL
Taq .2μL .2μL .2μL
H2O 3.8 3.8 3.8
Total 20μL 20μL 20μL

Expectatives

  • See in gel amplification of 3 extraction and not amplification in controls.


Personal tools