Automation of library preparation using Illumina ForenSeq kit for routine sequencing of casework samples

Abstract

Massively Parallel Sequencing (MPS) technologies are called upon to play a major role in forensic genomics in the next few years. However, library preparation protocols contain numerous steps which can complexify casework analysis in routine. Implementation of a MPS analytical workflow, in a casework laboratory, requires automation to guarantee a full sample tracking and to minimize contamination risks. In this study, we present the development and validation of a fully automated workflow using Illumina ForenSeq DNA signature Prep Kit on a NGS STARlet HID Illumina platform. Automation greatly helps with library preparation reproducibility and significantly reduces hands-on time. We are confident that automation of MPS technologies will enhance forensic laboratory to adopt these powerful analytical tools determinant for the future of the forensic genomics field.

Automation Management Training For Community Library

“…As the result, searching for the collection needed will be easier. The automation concept has created a more efficient way to organize the lending service (Laurent, 2017). The aims of the training are not only about introducing the automation community library program but also encouraging the library staff to develop the more efficient service to attract the visitors' attention to read.…”

Massive parallel sequencing in forensics: advantages, issues, technicalities, and prospects

“…One notable advantage of the Ion Torrent platforms is the availability of automated library preparation and chip loading stations, which simplifies the workflow considerably. Recently, similar automation and liquid handling solutions have been proposed for many MiSeq workflows [10].…”

A review of the method and validation of the MiSeq FGx™ Forensic Genomics Solution

“…During PCR2 contamination can occur if an adapter is added to the incorrect well on the PCR plate, or DNA transfers from one well to another. Laurent et al (2017) noted that contamination can only be detected if it occurs before or during the enrich targets step, as after this step each library has a sample specific index combination.…”

“…The beads have a fixed capacity so the same molarity of fragments will be retained. This method works well with libraries from DNA samples of 1 ng of input DNA but when libraries are made from lower concentration samples or from suboptimal quality samples, the number of reads produced decreases and becomes more variable (Churchill et al, 2016;Fattorini et al, 2017;Guo et al, 2017;Laurent et al, 2017;Moreno, Galusha, & Just, 2018). An alternative approach proposed by Hollard et al (2019) includes adjusting bead normalized libraries based on the amount of input DNA used, which produced consistent numbers of sequencing reads for libraries made from between 2 ng and 50 pg of input DNA.…”

“…A number of groups have developed methods to automate the ForenSeq™ library preparation. Laurent et al (2017) identified critical steps during the ForenSeq™ library preparation which are prone to the introduction of human errors and/or contamination. Using the Hamilton STARlet HID Illumina platform liquid handling robot, they developed methods to automate PCR1, PCR2, library purification, bead normalization, and library pooling steps, reducing hands on time to only 15 min, reducing the chance of contamination and human error.…”