Dna sequencing

DNA sequencing

The extraction of genomic DNA from the muscle of pereopods 3–7 from one paratype followed Tomikawa (2014). The cytochrome c oxidase subunit I (COI) gene [LCO1490 and HCO2198 ( Folmer et al.1994)] primer used for polymerase chain reaction (PCR) and cycle sequencing reactions. PCR reactions and DNA were performed following Tomikawa et al. (2016): the PCR mixtures were heated to 94°C for 7 min, followed cycles at 94°C (45 s), 42°C (1 min) and 72°C (1 min), and a final extension at 72°C (7 min); the cycle conditions were 25 cycles at 96°C (10 s), 50°C (5 s), and 60°C (4 min). DNA sequences have been deposited International Nucleotide Sequence Database Collaboration (INSDC) through the DNA Data Bank of Japan