Automated NGS Library Prep
Next Generation Sequencing (NGS) library preparation is a critical step in the sequencing workflow, involving the processing of DNA or RNA samples into a format that is compatible with NGS platforms.
This also includes bead clean up, which can be used for PC product isolation, or fragment purification for other applications outside of NGS.
Capable of handling up to 24 libraries, Myra incorporates a 96 Well Magnetic Station for bead clean-up steps, with off-deck heating to reduce robotic downtime. The workflow is split into manageable runs, with safe stopping points after the various library preparation steps including bead clean up.
Myra’s workflow is split into manageable runs, with safe stopping points after the various library preparation steps including bead clean up.
In NGS, repeatability and reproducibility are paramount. The Myra Liquid Handling System can successfully generate high-quality DNA libraries, with performance comparable to manual methods. Sequencing runs show high Q30 scores and consistent barcode balance. Sequencing coverage exhibits uniformity across the prepared libraries, resulting in low library variability and reliable performance. Thus, Myra not only reduces manual labour but also ensures high-quality sequencing results.
NGS Library Prep Kits on Myra
Browse a comprehensive table of all available templates on Myra, complete with direct links to specific MyraScript pages. Access protocol documentation via a dedicated PDF link. Protocols have been verified by BMS, Partner (kit company), and/or the Myra community (customers).
| Kit | Verified By |
|---|---|
| BMSCommunity |
|
| BMSCommunity |
|
| BMS |
|
| BMS |
|
| BMSCommunity |
Case Study: Illumina DNA Prep Kit
We validated up to 24 samples using the Illumina DNA Prep kit. Libraries displayed low variability across replicates, even barcode balance, and high coverage of sequence alignments.
Experimental Setup
Up to 24 samples prepared.
To demonstrate capability, 8-sample and 24-sample plates of 150 ng of input DNA (48 kB genome Lambda DNA and Human Genomic DNA) were prepared on the Myra Liquid Handling System. Manual preparation of an 8-sample plate of 150 ng of input DNA (48 kB genome Lambda DNA) was also performed as a comparison method. The quality of these libraries were checked with BiOptic Qsep1-Plus bio-fragment analyser (Figure 1). The libraries were quantified using NEBNext Library Quant Kit for Illumina with reaction setup on the Myra Liquid Handling System and amplification on the Mic qPCR Cycler. Libraries were simultaneously normalised and pooled to 2 M using the Myra Liquid Handling System, then sequenced on a 2×150 bp flow cell on an iSeq 100 System for whole genome sequencing. Q30 and percentage PF were used to show the quality of the libraries. Sequencing data was demultiplexed using Illumina’s BaseSpace Sequence Hub and coverage maps of the Lambda genome were analysed using Geneious Prime 2022.2.2.
Results
High quality library perp.
We were able to successfully generate high-quality 8-plex and 24-plex DNA libraries on the Myra, with comparative performance to the manual method (Figure 2). Fragment analyzer results show consistent fragment sizes across all 24 DNA libraries (Figure 1). The quality of the sequencing runs were comparative with Q30 score > 90% for all methods. Barcode balance was also accurate, with variability less than the manual method. Sequencing coverage of the Lambda genome also showed consistent uniformity across 8-plex and 24-plex DNA prep libraries constructed on the Myra. The sequence data showed low sample variability and reliable uniformity for libraries prepared using Illumina DNA Prep.
Additional Accessories
The following accessories are recommended for achieving library prep on the Myra liquid handling system.
Myra 96 Well Magnetic Station
MYRA-MAGS96
Designed to provide automated magnet control for up to 96 x 0.2 mL tubes, including strips of 8, and 96 tapered well plates (including fully skirted, unskirted and semi-skirted plates). Magnets are applied to groups of 24 wells at a time. Made from aluminium.
Myra 96 Well Cooling Block
MYRA-COOL96
Designed to provide passive cooling for 96 x 0.2 mL tubes including strips of 8, and 96 tapered well plates (including fully skirted, unskirted and semi-skirted plates).