Gene | Primer name | Primer sequence (5'-3) | Size of product (bp) | PCR program | M-PCR volume (50 μL) |
---|---|---|---|---|---|
pap | pap3 pap4 | GCAACAGCAACGCTGGTTGCATCAT AGAGAGAGCCACTCTTATACGGACA | 336 | 1 cycle: 94 °C ------------ 1 min. 30 cycle: 94 °C ------------ 60 s 63 °C ------------ 30 s 72 °C ------------ 90 s 1 cycle: 72 °C ------------ 5 min | 5 μL PCR buffer 10X 1.25 mM Mgcl2 150 μM dNTP (Fermentas) 1 μM of each primers F & R 1.2 U Taq DNA polymerase (Fermentas) 3 μL DNA template |
Sfa | sfa1 sfa2 | CTCCGGAGAACTGGGTGCATCTTAC CGGAGGAGTAATTACAAACCTGGCA | 410 | ||
Afa | afa1 afa2 | GCTGGGCAGCAAACTGATAACTCTC CATCAAGCTGTTTGTTCGTCCGCCG | 750 | ||
fimH | FimH1 FimH2 | GAGAAGAGGTTTGATTTAACTTATTG AGAGCCGCTGTAGAACTGAGG | 559 | 1 cycle: 94 °C ------------ 3 min. 40 cycle: 94 °C ------------ 60 s 58 °C ------------ 70 s 72 °C ------------ 70 s 1 cycle: 72 °C ------------ 6 min | 5 μL PCR buffer 10X 2 mM Mgcl2 200 μM dNTP (Fermentas) 0.4 μM of each primers F & R 1 U Taq DNA polymerase (Fermentas) 3 μL DNA template |