The application of a one-day hybridisation-based enrichment protocol for NGS incorporating a rapid (30 minute) hybridisation step

Friday 23 June 2017
Applications:

Lyudmila Georgieva, Ezam Uddin, Jacqueline Chan, Faidra Partheniou, and Graham Speight

Presented at the 42nd Association of Genetic Technologists (AGT) 2017 annual meeting in St Louis, USA, this poster outlines how OGT has optimised a one-day hybridisation-based enrichment protocol for next-generation sequencing (NGS) incorporating a rapid 30 hybridisation step, with performance examples illustrating excellent uniformity of coverage in the detection of key CALR and JAK2 indels (including 52 bp deletions and 5 bp insertions) and FLT3 ITDs.

Introduction

  • Hybridisation-based enrichment protocols for next-generation sequencing (NGS) generate higher quality data (e.g. enhanced coverage uniformity, more complete coverage, and more accurate assessment of insertions/deletions (indels) and internal tandem duplications (ITDs)). However, they are generally more time consuming than PCR-based enrichment approaches. 
  • We have developed a rapid (30 minute) hybridisation protocol that enables Illumina sequencer-ready libraries to be generated from purified DNA in 1-day. 
  • The aim of this study is to evaluate the new streamlined 1-day hybridisation-based NGS library preparation kit (LPK) in conjunction with four different haematological capture panels of varying sizes.

 

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