Design: Adult individuals were used for DNA extraction. Only the heads were used to minimize DNA contamination by the symbionts B. aphidicola and S. symbiotica whose genomes were sequenced previously. Whole insects were first surface sterilized with 99% ethanol, rinsed with sterile water and then immersed in 70% ethanol where the heads were dissected with microscissors before being stored directly in a sterile plastic tube at -80C prior to DNA extraction. DNA extraction was performed using a phenol-chloroform approach. Tissues were homogenized in STE buffer (100 mM NaCl, 1 mM Na2 EDTA, pH 7.8, 10 mM Tris-HCl, pH 8.0) with a sterile pestle, then treated successively by SDS 10%, proteinase K, and RNase. Briefly, genomic DNA was purified by two successive extractions with phenol:chloroform:isoamyl alcohol (25:24:1, v/v/v) followed by extraction with 1 vol of chloroform:isoamyl alcohol (24:1, v/v/v). Genomic DNA was then precipitated by 0.7 vol isopropanol. After washing the pellet with 70% ethanol, genomic DNA was recovered in TE buffer (1mM EDTA, 10mM Tris HCl pH 8). DNA concentrations and quality were assessed using NanoDrop (Thermo Fisher Scientific, Waltham, MA, United States), agarose gel electrophoresis and DNA concentrations and quality were measured using NanoDrop (Thermo Fisher Scientific, Waltham, MA, United States) and Qubit fluorometer (Thermo Fisher Scientific, Waltham, MA, United States). Sequencing libraries were prepared using the Express Template Prep kit 3.0 (Pacific Bioscience, Menlo Park, USA) and whole-genome sequencing was performed on the PacBio sequel IIe at the Genomics Core Leuven (KU Leuven, Leuven, Belgium) using the Sequel II Binding Kit 3.2 (Pacific Bioscience, Menlo Park, USA).
Submitted by: BF2I
Study:
Sipha maydis breed:Midelt Genome sequencingshow Abstracthide AbstractThis study aims at sequence and assemble the Sipha maydis genome
Library:
Name: bc2042
Instrument: Sequel II
Strategy: WGS
Source: GENOMIC
Selection: RANDOM
Layout: SINGLE
Runs:
1 run, 536,705 spots, 3.7G bases, 1.5Gb