10x genomic data of Xiphophorus variatus: from caudal fin tissue
Briefly, approximately 500 ng of DNA was sheared to ~400 basepairs using a Covaris sonicator. To repair the sheared ends, DNA was mixed with dNTPs and T4 DNA polymerase, Klenow DNA polymerase and T4 PNK and incubated at 25 oC for 30 minutes. After the end-repair reaction, the DNA was purified using 18% SPRI beads. A-tails were added by mixing the purified repaired DNA with dATPs and Klenow exonuclease and incubating at 37 oC for 30 minutes. After adding the A-tails, the DNA was purified using 18% SPRI beads and an adapter ligation reaction was performed. Purified DNA was amplified using indexed primers in an individual Phusion PCR reaction; between 11-15 PCR cycles were used. After amplification, libraries were purified using 18% SPRI beads. Libraries were quantified using a Qubit fluorimeter and library size distribution and quality were visualized on an Agilent Tapestation (Agilent, Santa Clara, CA). Libraries were sequenced on the Illumina NextSeq 4000 across four lanes to collect paired-end 75 basepair reads.