User:Jorge E. Buendia Buendia/Notebook/iGEM UNAM-Genomics-Mexico/2010/05/28

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May 28th,2010

1. PCR for Blue promoter and OGR with Paz’s reactives, control with primers for 1kb of plasmid with RFP (one reaction with Paz’s reactives and other with mines) and one positive control (I only added 1ul of Taq pol). I realized what was the mistake in the previous PCR’s, I used to add only Taq Polymerase Buffer but not Taq Polymerase or only Taq Pol and no Buffer, the new calculations are as follows for 1 reaction with 2ul of DNA:

  • Taq Polymerase 1
  • Taq Reaction Buffer 5
  • MgCl 50mM (can be used up to 3ul) 2.5
  • dNTP’s 0.4ug/ul 2.5
  • Primer Forward (can be used up to 3ul) 2.5
  • Primer Reverse (can be used up to 3ul) 2.5
  • HPLC 32
  • DNA 2
  • Total volume 50