With the advent of Next-Generation Sequencing (NGS), more and more researchers are turning to sequencing to better understand genetics and gene expression. It often seems like a daunting task to trust your precious rare samples for analysis on an instrument you don’t have much experience using.
Our SeqitOut video series tackles all things sequencing (from NGS to Sanger). To help you get started on your NGS experiments we’ve put together five tools to help you get started with your NGS experiments. And if a question pops into your head while you’re watching the videos, just send them in at thermofisher.com/ask – the team at Thermo Fisher Scientific just might make an episode out of it!
Let’s get started…
We all started with our ABC’s and somehow we ended up falling in love with AGTC’s. But when we step into NGS….OMG. Most of you already know the acronyms DNA and RNA. But, in the NGS world, there is a whole lot more. When you are starting to think about your next sequencing experiment, people may ask if you are doing WES, WGS, or target-seq? Let’s dive into the genetic dictionary that is Next-generation sequencing terminology.
The unprecedented high throughput of NGS instruments is helping drive down the cost of sequencing, making it accessible to large commercial and small research labs, alike. One way to take full advantage of high throughput NGS is sample multiplexing. Multiplex sequencing allows users to sequence more than one sample on the same sequencing run, reducing your cost per sample. Well, this sounds great. But, you may be asking yourself, how does it work? How many samples can I fit on my sequencing run?
A key step in the NGS workflow is preparing the input for sequencing, known as creating a library. An NGS library is a collection of similarly sized DNA fragments with known adapter sequences added to the 5’ and 3’ ends. A library corresponds to a single sample and multiple libraries, each with their own unique adapter sequences, can be pooled and sequenced in the same sequencing run. Here we focus on the four basic steps of NGS library preparation that can be broadly applied across different preparation methods.
Do you perform qPCR as a routine part of your research? Are you interested in gene expression, but haven’t yet looked into how next generation sequencing or NGS can help you out? Next generation sequencing, also known as NGS, provides opportunities to profile gene expression in a wide range of settings. Whether you are looking to discover potentially novel genes or transcripts, or study how certain genes respond to stimuli, NGS can offer a great complement to your existing methodology.
So you are seeing some variants in your NGS data that you are not confident about. How can you quickly re-sequence a particular locus of interest?
Have more questions? We’ll help you get them answered! Submit your questions at thermofisher.com/ask