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Table 1 List of reagents and PCR steps to amplify chloroplast regions trnL-F, rps16, ndhF and nuclear regions ETS and ITS2

From: Optimization of DNA extraction and PCR protocols for phylogenetic analysis in Schinopsis spp. and related Anacardiaceae

PCR reagents Final concentration
Chloroplast regions Nuclear regions
H2O s&d
Buffer 5× (Promega)
DMSO 1 M (Sigma-Aldrich) 10 %
BSA 400 ng/μl (Promega) 1/1000 1/1000
Cl2Mg (Promega) 5 mM 5 mM
Primer F 1 μM 1 μM
Primer R 1 μM 1 μM
dNTP’s 1 mM (Promega) 200 μM 200 μM
DNA dil 1/10 25 ng 25 ng
Taq pol 5u/μl (Promega) 1.5–2 U 1.5–2 U
PCR steps trnL-F/rps16 ndhF ETS ITS2
1—Initialization 2 min—97 °C 3 min—94 °C 10 min—95 °C 50 s—97 °C
2—Denaturation 1 min—94 °C 30 s—94 °C 1 min—94 °C 50 s—97 °C
3—Annealing 2 min—48/55 °C 1 min—48 °C 2 min—56 °C 50 s—53 °C
4—Extension 2 min—72 °C 50 s—72 °C 2 min—72 °C 1.5 min—72 °C
5—Elongation 16 min—72 °C 6 min—72 °C 16 min—72 °C 7 min—72 °C
  [steps 2–3–4] × 30 [steps 2–3–4] × 35 [steps 2–3–4] × 35 [steps 2–3–4] × 35