Table 3.
Targeted regions, primer sequences and approximate product length (bp) of PCR for primer pairs used to amplify six cpDNA regions of Norway spruce
| Region (short name) | cpDNA region | Primer | Sequence (5' - 3') | Position in cpDNA of Pinus thunbergii NC_001631 | Position in cpDNA of Picea sitchensis NC_011152 | Fragment Length in P. abies (bp) |
|---|---|---|---|---|---|---|
| ndhCp-ndhKp (CK) | pseudogene | ndhK-for1 | CACTTCAGTTCTTGTTGTTCC | 66530→66550 | 66774→66794 | 136 bp |
| ndhC_ndhK-rev | TCGCCACAGAACCAACGATG | 66666→66647 | 66910→66891 | |||
| psbM-trnD (MD) | intergene spacer | psbM_trnD-f1 | GTTCGAGTAACGGAATCTAAC | 28004→28024 | 27925→27945 | 200 bp |
| psbM_trnD-rev1 | CGAACGTCTTCTGGAGTAGC | 28204→28185 | 28124→28105 | |||
| Pt3024, ORF46b* (B) | microsatellite | B-for | GCTTATGGCATTGTTGATGT | 30166→30185 | 30701→30720 | 212-216 bp |
| B-rev | TGGGCATTCTAGCTGTATTG | 30387→30368 | 30941→30922 | |||
| Pt15169, ORF84* (D) | microsatellite | D-for | CTTGGATGGAATAGCAGCC | 15169→15187 | 15208→15226 | 124 bp |
| D-rev | GGAAGCGCATTAAGGTCATTA | 15286→15266 | 15327→15307 | |||
| trnT-trnL (TL) | intergene spacer | trnT_trnL-sp-f1 | CTGAGCTAAGCAGGCTCAATGG | 69189→69168 | 69573→69552 | 251 bp |
| trnT_trnL-sp-brev | GCATGTTATTATCCTCCCCTAG | 68945→68967 | 69323→69344 | |||
| trnL (Li) | Intron | trnL-intron-f2 | GAACGCTCTATTTACACC | 68497→68480 | 68884→68867 | 237 bp |
| trnL-intron-rev | ACACGTAGAATTGGACTCTATC | 68261→68282 | 68648→68669 | |||
| trnL-trnF (LF) | Intergene spacer | Aa_trnLF-for | GGTTCAAGTCCCTCTATCCC | 68198→68179 | ? | 186 bp |
| Aa_trnLF-rev | ACTGATCAACTCAACTTGTCATAAGATGG | 68014→68042 | 68400→68428 | |||
| matK-trnK (K2i) | intron | trnK2i-f1 | GCCCTCGTTCATGAGAATAACC | 3281→3302 | 3368→3389 | 204 bp |
| trnK2i-r1 | CATGAGTCAGGAGAGCGATTGG | 3477→3456 | 3573→3552 |
* cpDNA region names from Parducci et al. (2005); primers B and D are from Parducci et al. (2005); primers CK, MD, TL, Li, LF and K2i were designed specifically for this study