Table 3 |
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|
Primers and amplification conditions used in the study |
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| Forward primer (5'-3') |
Reverse primer (5'-3') |
Specific for |
Annealing temperature/time |
MgCl2 concentration (mmol/l) |
Size of product (bp) |
|
|
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| AGTAGTTTACTACTTTGCCG |
ACTGCTGCCTCCCGTAGGAG |
Universal (R1 and R2) |
58°C/60 secs |
1.5 |
350 |
| CATAACGTCGCAAGACCAAA |
GTGCAATATTCCCCACTGCT |
E. coli |
58°C/45 secs |
1.5 |
187 |
| TTGGGAATAACGGTTGGAAA |
TGTCTCAGTCCCAGTGTTGG |
Chlamydia spp. |
59°C/30 secs |
1.5 |
203 |
| CGCACGGGTGAGTAAGGTA |
GCGTCATAGCCTTGGTAAGC |
Campylobacter spp. |
66°C/1 min |
2.5 |
170 |
| AGTAGTTTACTACTTTGCCG |
CCGATGGCGTGAGGCCCTAA |
Yersinia spp. |
65°C/30 secs |
3.5 |
154 |
| CGCACGGGTGAGTAAGGTA |
GCTTAACACAAGTTGACTAG |
Campylobacterjejuni |
63°C/30 secs |
2.5 |
70 |
| CGCACGGGTGAGTAAGGTA |
GTCTTACATAAGTTAGATA |
Campylobacter concisus |
55°C/30 secs |
2.0 |
70 |
| CGCACGGGTGAGTAAGGTA |
ATACCTCATACTCCTATTTAAC |
Bacteroides ureolyticus |
55°C/30 secs |
2.0 |
70 |
|
|
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|
Specific oligonucelotides were designed as described here. Standard reaction mix was used with each set of oligonucleotides (see Materials and method), although annealing conditions varied, as did MgCl2 concentration. Denaturation conditions were always 94°C for 1 min and extension was performed at 72°C for 1 min (10 min final extension). Typically 35–40 cycles were performed. If no PCR product was detected, then a second round of amplification was performed using 2 μl of PCR product as template. |
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|
Cox et al. Arthritis Res Ther 2003 5:R1 doi:10.1186/ar602 |
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