Friday, 05 Jul 2013

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7/05/13 Primer Design for kerA biobrick

  • Designed primers for directed mutagenesis of our kerA biobrick using [http://www.bioinformatics.org/primerx/ PrimerX]:
  1. Primer pair 1 (EcoRI mutation, aat --> aac, asparagine)
    a. Forward: 5' GGTTAAAGTACTGAACTCAAGCGGAAGCGG 3'
    b. Reverse: 5' CCGCTTCCGCTTGAGTTCAGTACTTTAACC 3'
  2. Primer pair 2 (Pst1 mutation, gca-->gcg, alanine)
    a. Forward: 5' GTTGTCGTTGTAGCTGCGGCAGGGAACAGCGGATC 3'
    b. Reverse: 5' GATCCGCTGTTCCCTGCCGCAGCTACAACGACAAC 3'
  • Used APE for checking codons and mutating restriction sites
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