User:Jose Fabricio Lopez/Notebook/Logbook/2011/08/23

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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.

Gel.InsertoG.Back.pstb1t3.Banda.JPG

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.