NGS Library Preparation

Low-volume dispensing and miniaturization of library preparation for next-generation sequencing.

Introduction

Next-generation sequencing (NGS), a high-throughput method of single-cell sequencing, is compatible with a variety of samples, including microbial genomes and transcriptomes. Prior to sequencing, the genomes are fragmented into smaller sizes, and adapters are introduced for flow cell attachment during sequencing. These adapters can contain barcodes or indexes for multiplexing. NGS library prep of samples is an important final step before sequencing.

As NGS library prep is a significant cost of the total workflow, miniaturization of library prep enables the generation of high-quality samples while reducing overall costs. Cost also becomes more critical when many libraries are sequenced simultaneously. The I.DOT provides easy reaction miniaturization, while maintaining flexibility to perform various types of NGS library protocols.

Normalization

Indexing

Purification

Sequencing

How the I.DOT facilitates NGS library prep

Low volume non-contact dispensing.

Dispendix’s portfolio features the Immediate Drop-on-demand Technology (I.DOT), a patented approach for optimizing non-contact liquid-handling tasks. Specifically, the I.DOT provides easy reaction miniaturization, while maintaining flexibility to perform various types of NGS library protocols. The system uses eight individually controlled positive pressure channels to generate droplets from 8 to 50 nL from a small outlet at the bottom of each well. Each channel can generate up to 100 droplets per second, which enables a volume range from 8 nL to 80 μL. The non-contact dispensing significantly reduces the number of pipette tips required during NGS library prep, and the low-volume dispensing (dead volume <1 µL) provides significant cost savings on reagents.

Quantification and Normalization of NGS libraries

Typically, complementary DNA (cDNA) amplification from single cells results in uneven concentrations, which may require normalization to ensure an even read distribution for all cells. Hundreds of libraries can be quantified and normalized within minutes using the I.DOT, which allows for highly multiplexed dispensing of up to 96 reagents in parallel.

LIBRARY INDEXING AND EARLY BARCODING

The ability to index NGS libraries with unique DNA barcodes provides the basis for high-throughput single-cell sequencing. For single-cell full-length transcript sequencing or whole exome sequencing, individual libraries are usually indexed after amplification and fragmentation.

PERFORMING COMPLEX TASKS AT UNRIVALED SPEED

The I.DOT non-contact dispensing technology was developed to perform even the most complex liquid-handling tasks at unrivaled speeds. Up to 2304 libraries can be uniquely barcoded with a single I.DOT source plate. Dispensing 96 unique barcodes takes less than 60 seconds.

Related products

I.DOT

A premier non-contact liquid-handling solution designed to provide efficiency and speed to your dispensing processes. The I.DOT can assist in miniaturizing your assays, compound screening or sample prep.

Dispensing plates

High quality dispensing plates, configured to meet your low- and high-throughput needs.

C.WASH™

Automated, rapid, and highly reproducible washing of entire 96-, 384-, and 1536-well plates.

plexWell™ Rapid Single Cell kit

Premier single cell RNA Sequencing.

plexWell™ Library Prep kit

Simple and scalable multiplexed workflow for 1000’s of samples.