Streptococcus oralis MLST primers and PCR conditions

Primers

Gene Primer name Sequence
Shikimate dehydrogenase aroE-F 5'-GCCTTTGAGGCGACAGC-3'
aroE-R 5'-TGCAGTTCA(G/A)AAACAT(A/T)TTCTAA-3'
D-alanine-D-alanine ligase ddl-F 5'-TGC(C/T)CAAGTTCCTTATGTGG-3'
ddl-R 5'-CACTGGGT(G/A)AAACC(A/T)GGCAT-3'
Glucose-6 phosphate dehydrogenase gdh-F 5'-ATGGACAAACCAGC(G/A/T/C)AG(C/T)TT-3'
gdh-R 5'-GCTTGAGGTCCCAT(G/A)CT(G/A/T/C)CC-3'
Glucokinase gki-F 5'-GGCATTGGAATGGGATCACC-3'
gki-R 5'-TCTCCCGCAGCTGACAC-3'
Mismatch repair protein hexB-F 5'-ACCTATCTCTTTGCCCAG-3'
hexB-R 5'-CCTGAATACGTCGGAACATCTTT-3'
Transketolase recP-F 5'-GCCAACTCAGGTCATCCAGG-3'
recP-R 5'- TGCAACCGTAGCATTGTAAC-3'
Xanthine phosphoribosyltransferase xpt-F 5'-TTATTAGAAGAGCGCATCCT-3'
xpt-R 5'-AGATCTGCCTCCTTAAATAC-3'

 

Thermocycling conditions for PCR

  • Initial denaturation: 96°C for 5 minutes.
  • 30 cycles of 94°C for 30 seconds, 49°C for 30 seconds and 72°C for 90 seconds.
  • Final extension at 72°C for 5 minutes.