Friday, 05 Jul 2013

From 2013.igem.org

7/05/13 Primer Design for kerA biobrick

• Designed primers for directed mutagenesis of our kerA biobrick using 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