2.2 DNA extraction and sequencing
High-quality genomic DNA was extracted from blood samples using a
modified CTAB (Hexadecyl Trimethyl Ammonium Bromide) method. The quality
and quantity of the extracted DNA were examined using a NanoDrop 2000
spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA), Qubit ds
DNA HS Assay Kit on a Qubit 3.0 Fluorometer (Life Technologies,
Carlsbad, CA, USA), and electrophoresis on a 0.8% agarose gel.
A paired-end sequencing library with an insertion length of 250 bp was
constructed using the VAHTS Universal DNA Library Prep Kit for MGI
(Vazyme, Nanjing, China). The Agilent 2100 Bioanalyzer (Agilent
Technologies, Santa Clara, CA) was used to validated the purity and size
distribution. Then, the obtained library was sequenced with the
paired-end, 150-bp mode using the MGI-SEQ2000 platform by Frasergen
Bioinformatics Co., Ltd. (Wuhan, China).
Ten micrograms (ug) of C. sonnerati genomic DNA were used for 20
kb template library preparation using the BluePippin Size Selection
system (Sage Science, USA) following the manufacturer’s protocol. The
library was sequenced on the Pacfic Biosciences Sequel II platform.