Next-generation sequencing (NGS) and single-cell technology share many overlaps, which is perhaps why they’re often confused. However, both terms sit in their own respective corners of the pentacontagon of biological understanding—that’s a 50-sided polygon, for reference.
In the same way that two books (one science fiction and one high fantasy) might be categorized next to each other in the fiction section of a library, NGS and single-cell technology contain their own world of research and understanding.
However, one easy distinction is that single-cell genomics & technology involves the whole cell, whereas next-generation sequencing only involves one piece of the cell (DNA or RNA sequencing analysis). This is, again, too simple of analysis to be accurate, but it will do as a brief introduction to the two.
To completely understand the difference, you need to isolate each concept. Only then will they stand out as their own pockets of research. Thus, this guide will be separated into two parts:
By the end, you should have a clear picture of both concepts, their common techniques and technologies associated, and the analytical gap between them.
NGS is the overarching term that encompasses the techniques and technologies available today that allow researchers to sequence strands of DNA and RNA. This allows them to study individual sections, or genes, and identify which sections of genomes contribute to disease or biological characteristics.
How is this accomplished? And what is the specific sequencing technology and method involved?
Regardless of instrumentation (NGS technologies), next-generation sequencing follows a similar workflow (NGS techniques) every time. There are four steps to the NGS workflow:
To gain more insight into the NGS workflow and processes, you need to know a bit about the actual methods involved.
Related: RNA Sequencing Analysis
Next-generation sequencing involves common technologies that have been used for the last 15 to 20 years. These are known as “third generation” sequencing methods—the second generation and first generation of these sequencing techniques, including Sanger sequencing, takes scientists back to the 1970s.
What are some next-generation sequencing methods?
If there is already genomic information available about what you’re studying, you can often use two methods that work on targeted sequences:
By now, you (hopefully) have a solid understanding of Next-Generation Sequencing and are ready to understand the other concept we have on our plate. Single cell technology.
As a fundamental understanding, single cell technologies are tools within cell biology and microbiology to analyze cells and cell populations. These tools are incredibly useful when most samples that researchers have to work with come in heterogeneous populations.
Single-cell methods, such as DNA sequencing technology, are then the complementing tools to bulk analysis methods (which have long been in use).
In a 2016 issue of Nature surrounding single cell technology, they present their focus piece to describe the rising importance of single cell technology in unmasking the inherent heterogeneity of various sample tissues:
“With the increasing sophistication of microfluidics, electrophysiology measurements, high-resolution imaging, deep sequencing and mass spectrometry platforms, a more detailed picture of cellular subtype, physical location in tissue, and clonal evolution is emerging. Moreover, the exquisite sensitivity of these approaches is enabling the identification of rare cells of potential functional or pathogenic consequence.”
This offers a succinct way to describe the well of information needed to inform single cell technology.
From there, the specific technologies and methodologies will inform much about what this branch of cell biology covers.
Single-cell DNA sequencing is considered one of the many single-cell technologies. So, while there are many differences when talking about both terms, now you can glean some insight into how they are similar.
Let’s discuss the other single-cell technologies.
It’s much easier to work on single-cell analysis with a homogenous population. Flow cytometers help with this.
Cytometry is a method that sorts cells based on certain characteristics in a highly efficient manner. Using properties of fluid dynamics, a cytometer is able to create a flow that allows cells within a given sample to pass through a laser one at a time (with very little couplets or error).
From there, laser optics and measuring devices are used to identify the cell, and electronics systems charge the cell either positive or negative (or remain neutral), which then isolates cells into one of three categories.
This is a primary resource in nearly all cell research labs, with the only downside being the high pressures can cause abrasion and cell death in the population.
That is, until NanoCellect created the WOLF G2 Cell Sorter. By using sorting pressures of <2 psi, it is officially 30 times gentler than industry-standard flow cytometers. Additionally, the WOLF Cell Sorter is:
Once the cells are sorted, the process of cloning allows you to create a large homogenous sample from just a few individual cells. The techniques involved include:
RT-PCR allows researchers to quickly and efficiently multiply the amount of DNA strands available for analysis. This can be done on the scale of millions or billions of copies, depending on what’s needed.
The steps for RT-PCR include:
You can probably see where a lot of the overlap comes into play between next-generation sequencing and single-cell technology. Both techniques are sensitive to contamination and, despite the major advances in the field, there is still room for error. Thus, single-cell and NGS technology both rely on precise, gentle handling.
This is where having the proper cell sorter comes in. NanoCellect’s WOLF Cell Sorter enables healthy, viable cells. This allows NGS, single-cell multiomics and antibody development to be done efficiently.
That’s one more thing they have in common.
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