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Establishing the limit of detection of massively parallel sequencing using laser micro-dissected cells
online resourceposted on 09.04.2019, 02:05 authored by Gemma NancollisGemma Nancollis, Ryan England, SallyAnn Harbison
Massively parallel sequencing is fast emerging as an increasingly useful tool for forensic science. As part of our ongoing validation of this technology we wanted to both ensure compatibility of the technology with laser microdissected (LMD) cells and use the LMD to explore the minimal and optimal numbers of cells required to generate informative profiles. To do this we used the ForenSeq (TM) DNA Signature Prep Kit and a MiSeq FGx (TM) Sequencer. In this paper we describe the modifications made to our one step DNA extraction protocol to optimize sequencing performance, and address technical issues that require consideration when sequencing DNA from small numbers of cells, below the optimum amounts recommended. We demonstrate that DNA sequence profiles can be obtained from very small numbers of laser micro dissected epithelial and sperm cells, demonstrating the utility of this approach.