Melbourne/GvpL site dirrected primers

From 2007.igem.org

Primer sequences: Primer Name Primer Sequence (5' to 3')

g213a 5'-aggatatgagctggctgcaagaaaaagcatttcatcatc-3'
g213a_antisense 5'-gatgatgaaatgctttttcttgcagccagctcatatcct-3'
g318a 5'-ttataaaggtgaagaaagtctgcaagcagctattgagattaacaaag-3'
g318a_antisense 5'-ctttgttaatctcaatagctgcttgcagactttcttcacctttataa-3'
g351a 5'-attgagattaacaaagaaaagatagaaaattcactgacgctgcttc-3'
g351a_antisense 5'-gaagcagcgtcagtgaattttctatcttttctttgttaatctcaat-3'
g696a 5'-ttcttctcccgtctctgcaaattactaagttcgtaaacg-3'
g696a_antisense 5'-cgtttacgaacttagtaatttgcagagacgggagaagaa-3'

Oligonucleotide information: Tm and CG content verified from http://mbcf.dfci.harvard.edu/docs/oligocalc.html

Primer Name Length (nt.) Tm Duplex Energy at 68°C Energy Cost of Mismatches GC content Tm verOD=1bp beforebp after
g213a 39 78.45°C -44.89 kcal/mole 6.5% 41% 81 deg C 2216nM1919
g213a_antisense 39 78.45°C -42.10 kcal/mole7.8% 41% 32% 2216nM
g318a 47 78.97°C -46.02 kcal/mole 6.3% 30% 80 deg C 1754nM2521
g318a_antisense 47 78.97°C -42.40 kcal/mole 7.7% 30% 80 deg C 1754nM
g351a 46 78.02°C -43.40 kcal/mole 5.3% 33% 80 deg C1820nM2619
g351a_antisense 46 78.02°C -40.27 kcal/mole 7.4% 33% 80 deg C1820nM
g696a 39 78.45°C -42.46 kcal/mole 5.4% 41% 812496nM1919
g696a_antisense 39 78.45°C -43.97 kcal/mole 6.8% 41% 81 Deg C2496nM