Dna extraction

Khan, Nazar, Lyskov, Dmitry, Sultan, Amir, Ukrainskaja, Uliana, Samigullin, Tahir, Khan, Tahir, Ishaq, Kamran & Khan, Amjad, 2025, Semenovia zhobica (Apiaceae), a new species from Northern Balochistan, Pakistan, Phytotaxa 695 (1), pp. 91-104 : 92

DNA extraction , amplification and sequencing

Total genomic DNA was isolated from fruit material using a Diamond DNA Plant kit (ABT, Barnaul, Russia) following the manufacturer’s instructions. Polymerase chain reaction (PCR) amplification was performed on a Biometra T3000 Thermocycler using an Encyclo PCR kit (Evrogen JSC, Moscow, Russia). Details of the PCR for nrITS region amplifications (including primer locations and characteristics) and sequencing strategies for nrITS were described by Valiejo-Roman et al. (2002). Primers for sequencing the ETS fragment and conditions of its amplification and sequencing were described by Logacheva et al. (2010). Cycle sequencing was performed by automated sequencing using Big Dye Terminator version 3.1 for both forward and reverse strands on ABI Prism 3100-Avant (Applied Biosystems). Newly obtained sequences were deposited in the GenBank (Appendix 1).