User:Eduardo Vladimir Munoz/Notebook/UNAM Genomics Mexico 2011/2011/08/23

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High Stability Plasmid Development

Abstract

  • repC PCR cleanage
  • repC PCR digestion

PCR cleanage

Today I cleaned the repC region PCR that Fibo performed with Mac. It was diluted finally to 60μL.

PCR digestion

The repC region it was ampliffied with oligonucleotides containing the standard preffix and suffix. To assemble it with the plasmid containing a transcriptional terminator pSB1AK3, placing the terminator after the part, I digested with the enzymes EcoRI and SpeI:

repC PCR 15μL
BSA 1μL
Buffer 2 5μL
EcoRI 2μL
SpeI 2μL
H2O mQ 25μL
Total 50μL