Phylogeny and classification of the East Asian Amitostigma alliance (Orchidaceae: Orchideae) based on six DNA markers

Table 1 PCR cycling parameters for all DNA regions included in this study

DNA marker	nrITS		matK		trnL-F		trnS-trnG		psbA-trnH		Xdh
Initialization	94 °C	240 s	94 °C	240 s	94 °C	240 s	94 °C	240 s	94 °C	240 s	cf. Górniak et al. [48]
Denaturation	94 °C	40–50 s	94 °C	45 s	94 °C	40–50 s	94 °C	45 s	94 °C	45 s
Annealing^a	55 °C	40–50 s	48 °C	40–50 s	55 °C	40–50 s	52 °C	50–60 s	53 °C	50 s
Extension	72 °C	50–60 s	72 °C	60 s	72 °C	50–60 s	72 °C	50–60 s	72 °C	50–60 s
Number of cycles	30		30		30		30		30
Final extension	72 °C	420 s	72 °C	420 s	72 °C	420 s	72 °C	420 s	72 °C	420 s

^aInitially, the annealing temperatures for all regions were set to 53 °C; for samples with low yield, the temperatures were then adjusted to those specific for each region

ISSN: 2730-7182