Complete Genome Sequence of Lumpy Skin Disease Virus isolated from Rangpur, Bangladesh.
Shotgun metagenomic libraries were prepared following a modified iNextEra protocol. Briefly, DNA was tagmented using Illumina bead-linked transposomes (Illumina DNA Library Prep, Illumina, Inc.) and 2X TMP buffer in a 6 L reaction (2.5 L DNA, 3 L 2X TMP, 0.5 L transposome) at 53C for 30 min. Tagmented DNA was amplified with limited-cycle PCR to add dual indexes using custom iNextEra primers. Each 62.5 L PCR contained 51.5 L master mix (HyClone water, Q5 buffer, dNTPs, and Q5 polymerase), 6 L tagmentation mix, and 5 L index primers. Cycling conditions were: 72C for 3 min, 98C for 3 min, 12 cycles of 98C for 45 s, 62C for 30 s, 72C for 2 min, and a final extension at 72C for 1 min. PCR products (300700 bp) were confirmed by agarose gel, quantified with Qubit Flex, normalized, pooled, and purified with AMPure XP beads. The pooled libraries were sequenced on an Illumina NovaSeq X Plus platform (2150 bp, Novogene, USA).
SUB15965880
LSDV_Rangpur-05-MGH-BD