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Table 1 The oligonucleotide primers and the Multiplex PCR programs used for amplification of O-serogroups genes of E. coli isolates

From: Biofilm formation, antimicrobial susceptibility, serogroups and virulence genes of uropathogenic E. coli isolated from clinical samples in Iran

Serotypes

Gene

Primer name

Primer Sequence (5'-3)

Size of product (bp)

PCR programs

M-PCR Volume (50 μL)

O1

Wzx

wl-14632

GTGAGCAAAAGTGAAATAAGGAACG

1098

1 cycle:

95 °C ------------ 5 min.

30 cycle:

95 °C ------------ 30 s

62 °C ------------ 60 s

72 °C ------------ 60 s

1 cycle:

72 °C ------------ 5 min

5 μL PCR buffer 10X

2.5 mM Mgcl2

300 μM dNTP (Fermentas)

0.4 μM of each primers F & R

2 U Taq DNA polymerase (Fermentas)

3 μL DNA template

wl-14633

CGCTGATACGAATACCATCCTAC

O6

Wzy

wl-14646

GGATGACGATGTGATTTTGGCTAAC

783

wl-14647

TCTGGGTTTGCTGTGTATGAGGC

O7

Wzx

wl-14648

CTATCAAAATACCTCTGCTGGAATC

610

wl-14649

TGGCTTCGAGATTAAACCTATTCCT

O8

orf469

wl-14652

CCAGAGGCATAATCAGAAATAACAG

448

wl-14653

GCAGAGTTAGTCAACAAAAGGTCAG

O16

Wzx

wl-14654

GGTTTCAATCTCACAGCAACTCAG

302

wl-14655

GTTAGAGGGATAATAGCCAAGCGG

O21

Wzx

wl-14676

CTGCTGATGTCGCTATTATTGCTG

209

wl-14677

TGAAAAAAAGGGAAACAGAAGAGCC

O75

Wzy

wl-17413

GAGATATACATGGGGAGGTAGGCT

511

wl-17414

ACCCGATAATCATATTCTTCCCAAC

O2

Wzy

wl-14636

AGTGAGTTACTTTTTAGCGATGGAC

770

1 cycle:

95 °C ------------ 5 min.

30 cycle:

94 °C ------------ 60 s

58 °C ------------ 60 s

5 μL PCR buffer 10X

2.5 mM Mgcl2

300 μM dNTP (Fermentas)

0.4 μM of each primers F & R

2 U Taq DNA polymerase (Fermentas)

wl-14637

AGTTTAGTATGCCCCTGACTTTGAA

O4

Wzx

wl-14642

TTGTTGCGATAATGTGCATGTTCC

664

wl-14643

AATAATTTGCTATACCCACACCCTC

O15

Wzy

wl-14672

TCTTGTTAGAGTCATTGGTGTATCG

183

wl-14673

ATAAAACGAGCAAGCACCACACC

O18

Wzx

wl-14656

GTTCGGTGGTTGGATTACAGTTAG

551

wl-14657

CTACTATCATCCTCACTGACCACG

O22

Wzx

wl-14660

TTCATTGTCGCCACTACTTTCCG

468

wl-14661

GAAACAGCCCATGACATTACTACG

O25

Wzy

wl-14666

AGAGATCCGTCTTTTATTTGTTCGC

230

wl-14667

GTTCTGGATACCTAACGCAATACCC

O83

Wzx

wl-14668

GTACACCAGGCAAACCTCGAAAG

362

wl-14669

TTCTGTAAGCTAATGAATAGGCACC