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Table II.
PCR primers used for identification of isolates and SSCP analysis.
| Primers1 | Species target | Sequences | PCR conditions | Target |
|---|---|---|---|---|
| W02 | Bacterial 16S rDNA | GNTACCTTGTTACGACTT | 25 cycles of: 30 s at 94 °C, 30 s at 50 °C, 90 s at 72 °C | SSU 16S rDNA |
| W18 | GAGTTTGATCMTGGCTCAG | |||
| W34 | Partial bacterial 16S rDNA | TTACCGCGGCGTGCTGGCAC | 30 cycles of: 30 s at 96 °C, 30 s at 61 °C, 30 s at 72 °C | SSU 16S rDNA |
| W49 | ACGGTCCAGACTCCTACGGG | |||
| Gram1F | Partial bacterial 16S rDNA | CCTAATACATGCAAGTCG | 30 cycles of: 30 s at 96 °C, 30 s at 52 °C, 45 s at 72 °C | SSU 16S rDNA |
| Gram2R | CTCAGTCCCAATGTGGCC | |||
| nl1 | Yeast 26S rDNA | GCATATCAATAAGCGGAGGAAAAG | 30 cycles of: 30 s at 94 °C, 30 s at 57 °C, 30 s at 72 °C | 26S rDNA |
| nl4 | GGTCCGTGTTTCAAGACGG | |||
| Paracasei 16S | Lactobacillus casei/paracasei | CACCGAGATTCAACATGG | 30 cycles of: 60 s at 94 °C, 00 s at 53 °C, 60 s at 72 °C | 16S rDNA |
| 16rev | GAAAGGAGGTGATCCAGC | |||
| 16 forward | Lactobacillus plantarum | GCTGGATCACCTCCTTTC | 30 cycles of: 60 s at 94 °C, 00 s at 53 °C, 60 s at 72 °C | 16S/23S spacer region |
| Lpl | ATGAGGTATTCAACTTATT | |||
| 16 forward | Lactobacillus curvatus | GCTGGATCACCTCCTTTC | 30 cycles of: 60 s at 94 °C, 00 s at 53 °C, 60 s at 72 °C | 16S rRNA gene |
| Lc | TTGGTACTATTTAATTCTTAG | |||
| Lnm1 | Leuconostoc mesenteroides/pseudomesenteroides | TGTCGCATGACACAAAAGTTA | 25 cycles of: 30 s at 94 °C, 30 s at 62 °C, 90 s at 72 °C | 16S rDNA position 185 forward |
| Lnm2 | ATCATTTCCTATTCTAGCTG | 16S rDNA position 470 reverse | ||
| Lncit1 | Leuconostoc citreum | ACTTAGTATCGCATGATATC | 30 cycles of: 30 s at 94 °C, 30 s at 58 °C, 90 s at 72 °C | 16S rDNA position 183 forward |
| Lncit2 | AGTCGAGTTGCAGACTGCAG | 16S rDNA position 1326 reverse | ||
| ddlE1 | Enterococcus faecalis | ATCAAGTACAGTTAGTCT | 30 cycles of: 60 s at 94 °C, 60 s at 54 °C, 60 s at 72 °C | glycopeptide resistance gene |
| ddlE2 | ACGATTCAAAGCTAACTG | |||
| ddlF1 | Enterococcus faecium | TAGAGACATTGAATATGCC | 30 cycles of: 60 s at 94 °C, 60 s at 54 °C, 60 s at 72 °C | |
| ddlF2 | TCGAATGTGCTACAATC |