Massively parallel sequencing or next generation sequencing is becoming a ubiquitous technology in
basic biology research and starting to make inroads in both diagnostic and clinical settings.
Despite its popularity, understanding how it works usually requires reading several manuscripts,
translating technology jargon and stitching together tidbits from presentations or discussions with
colleagues. This handbook is designed to assist those new to next generation sequencing get from
nucleic acid sample to next generation sequencing data on the first try. Avoiding sequencing redo’s
saves a lot of time and research money!
Section I, Designing a Sequencing Run outlines what you should consider when you first decide you’d
like to sequence your sample. Section II, Library Preparation goes into choosing a library prep kit,
considerations for calculating the coverage you need for DNA or RNA applications and determining
whether targeted or global whole genome sequencing makes more sense for your application. Section
III, briefly summarizes considerations for how your DNA or RNA should be fragmented and best
practices for isolation and shipping your sample to a NGS services provider. Section IV,
Recommendations for Analyzing your Sequencing Data, highlights types of biotinformatics services and
the providers available to perform analysis of your raw sequencing data.
This is a living handbook, meaning we actively update it to keep current with fast changing
technology. If you’d like to add something, please send us a message at firstname.lastname@example.org.
I. Designing a Sequencing Run
II. Library Preparation
Library prep considerations for:
III. Preparing your Sample for Library Preparation
IV. Recommendations for Analyzing Your Sequencing Data