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.