“…To determine the nucleotide sequence of the structural polyprotein of CHIKV, total extracted RNA from real-time RT-PCR-positive samples was converted into cDNA using the superscript III first-strand synthesis system (Invitrogen, Carlsbad, CA, USA). Briefly, 4 µL RNA were mixed with a specific primer [47], dNTP, buffer, MgCl 2 , and DTT following the manufacturer's protocol, and then 2 µL of 3 half cDNA was further amplified with Primestar GXL DNA polymerase (Takara, Japan) using 3 primer sets [47,48], chf18/chr24, chf20/chr24, and chf23/3RT, to obtain 3 overlapping amplified products of 2.7 kb, 1.9 kb, and 2.0 kb. The amplicons were purified, cleaned (Nucleospin, MACHEREY-NAGEL, Germany), and sequenced (Macrogen, Seoul, Korea) using the primers chr20, chr21, chr22, chf21, chf22, chf24, and chf25 [48].…”