TABLE 2.
Primers used in this study
Primera | Sequence (5′-3′) | Specificity | Annealing temp (°C) | DGGE gradient (%)c | Reference |
---|---|---|---|---|---|
968f-GCb | AAC GCG AAG AAC CTT AC | Bacterial V6-V8 region | 56 | 30-55 | 40 |
1401r | CGG TGT GTA CAA GAC CC | Bacterial V6-V8 region | 56 | 40 | |
Bif164f | GGG TGG TAA TGC CGG ATG | Bifidobacterium | 66 | 45-55 | 22 |
Bif662r-GCb | CCA CCG TTA CAC CGG GAA | Bifidobacterium | 66 | 22 | |
Lac1f | AGC AGT AGG GAA TCT TCC A | LAB | 61 | 30-55 | 65 |
Lac2r-GCb | ATT YCA CCG CTA CAC ATG | LAB | 61 | 65 | |
g-Bfra-F-GCb | ATA GCC TTT CGA AAG RAA GAT | Bacteroides fragilis group | 50 | 30-50 | 33 |
g-Bfra-R | CCA GTA TCA ACT GCA ATT TTA | Bacteroides fragilis group | 50 | 33 | |
Sg-Clept-F | GCA CAA GCA GTG GAG T | Clostridium leptum subgroup | 50 | 30-50 | 34 |
sg-Clept-R3-GCb | CTT CCT CCG TTT TGT CAA | Clostridium leptum subgroup | 50 | 34 | |
g-Ccoc-F-GCb | AAA TGA CGG TAC CTG ACT AA | Clostridium coccoidessubgroup | 50 | 35-50 | 33 |
g-Ccoc-R | CTT TGA GTT TCA TTC TTG CGA A | Clostridium coccoides subgroup | 50 | 33 |
f (forward) and r (reverse) indicate the orientation of the primers with respect to the 16S rRNA gene sequence.
GC indicates a 40-bp GC-rich sequence attached to the 5′ end of the primer: 5′-CCCGCCGCGCCCCGCGCCCGTCCCGCCGCCCCCGCCCG-3′.
Denaturing acrylamide of 100% was defined as 7 M urea and 40% (vol/vol) formamide.