Sequencing of Yellow and Green Morphs

Tail-tip tissue samples of 30 yellow and 30 green morphs were collected, and high-quality DNA was extracted using a QIAGEN® Genomic kit. After monitoring degradation and contamination using 1% agarose gels, DNA purity was checked using a NanoPhotometer® spectrophotometer (IMPLEN, CA, USA). DNA concentration was measured using a Qubit® DNA Assay Kit in Qubit®2.0 Fluorometer (Life Technologies, CA, USA). In total, 700 ng of DNA per sample was used as input material for DNA sample preparation. Sequencing libraries were generated using a NEB Next® Ultra DNA Library Prep Kit for Illumina® (NEB, USA) following the manufacturer’s recommendations and index codes were added to attribute sequences to each sample. The PCR products were purified (AMPure XP system), and library quality was assessed on the Agilent Bioanalyzer 2100 system. The clustering of the index-coded samples was performed using cBot Cluster Generation System with a NovaSeq 6000 PE Cluster Kit (Illumina) according to the manufacturer’s instructions. After cluster generation, the prepared library was sequenced on the Illumina NovaSeq 6000 platform and 150bp paired-end reads were generated.