Covaris Shearing Guide for 500 Base Pairs using the 96 oneTUBE-10 AFA Plate
Library preparation for high-throughput whole-genome sequencing (WGS) requires an automation-ready workflow and larger insert sizes than targeted sequencing. The oneTUBE™-10 product line, developed to meet high-throughput demands, is designed to adhere to automation microtiter plate standards (ANSI/ SLAS 4-2004 (R2012)) and is available in an 8-strip or 96-well plate format. This application note provides additional support for the current oneTUBE-10 shearing protocol to achieve 500 base pair (bp) DNA fragments using Adaptive Focused Acoustics® (AFA®) on the LE220-plus Focused-ultrasonicator. This protocol is optimized for the 500 cycle Illumina® sequencing chemistry used in WGS and is also compatible with PCR-free library construction, including the NxSeq® AmpFREE Low DNA Library Kit (Lucigen®). Here, we present two variations of sample preparation techniques utilizing the Covaris truSHEAR buffer, which yield comparable, qualitative results. This protocol is ideal for high-throughput analyses and enables flexibility in sample elution methods.