Performing next-generation sequencing can be complex, particularly for newcomers, with challenges in sample preparation, data reliability and protocol adherence. These hurdles can lead to frustration and suboptimal results.
This guide provides readers with practical tips and detailed advice to help them overcome common obstacles and achieve success in their sequencing projects.
Download this guide to discover:
- How to select the right nucleic acid isolation kit to ensure sample quality
- Essential QC measures to maintain the integrity of your DNA and RNA samples
- Tips on choosing the best library preparation kits and sequencing configurations for your specific research needs
How To Guide
Next-Generation Sequencing:
Top Tips To Guarantee Success
Elsa Molina, PhD
Next-generation sequencing (NGS), also known as deep sequencing or second-generation sequencing,
is a high-throughput and rapid sequencing technology that allows you to analyze large amounts of DNA
or RNA in parallel.1 With the advent of NGS, the cost of sequencing has decreased massively, improving
accessibility and greatly accelerating progress in whole-genome sequencing, transcriptome analysis,
and de novo sequencing.
While sequencing has become much more affordable, navigating the entire process – from nucleic acid
isolation to the sequencing run – can be a challenging journey, especially for scientists who are new to the
field. The complexity of sequencing methods and sample preparation can raise many questions and lead to
confusion, potentially resulting in unreliable data, or data that cannot be analyzed effectively
This guide, focused on bulk sequencing, is designed to provide you with tips on how to conduct a successful
sequencing project, helping you gain a clear understanding of each step in the process.
A dedicated nucleic acid isolation kit is key
For bulk DNA or RNA sequencing, the process begins with the isolation of DNA or RNA. The choice of isola
tion
kit is key here. Choose a brand recognized for the quality of its nucleic acid products and ensure a good
quality sample, as this is an essential requirement for sequencing.
Select the appropriate isolation kit based on the following criteria to maximize nucleic acid quality and yield:
1. Type of tissue or samples
Isolating nucleic acids from some tissues or samples can be more challenging than others – for ex
ample,
isolating RNA from a tissue rich in RNases (e.g., pancreas), a fibrous tissue (e.g., muscle), a fatty
tissue (e.g., liver) or a calcified tissue (e.g., bone). In addition, isolating genomic DNA (gDNA) from soil
(e.g., sediment) can pose difficulties. It’s worth noting that there are typically dedicated kits available for
these troublesome tissues!
2. Species
Companies offer different categories of kit based on the type of animal (e.g., mammalian or non-mam
malian)
or even specific species. Make sure to select the most appropriate one for your project.
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