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Sanger Sequencing01:57

Sanger Sequencing

760.7K
DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...
760.7K
RNA-seq03:21

RNA-seq

10.5K
RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
10.5K
Maxam-Gilbert Sequencing01:05

Maxam-Gilbert Sequencing

11.6K
In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
Challenges of the Maxam-Gilbert Method
The...
11.6K
Next-generation Sequencing03:00

Next-generation Sequencing

93.7K
The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features....
93.7K
Peptide Identification Using Tandem Mass Spectrometry01:33

Peptide Identification Using Tandem Mass Spectrometry

7.2K
Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
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Related Experiment Video

Updated: Oct 7, 2025

Sequencing of mRNA from Whole Blood using Nanopore Sequencing
11:26

Sequencing of mRNA from Whole Blood using Nanopore Sequencing

Published on: June 3, 2019

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Protein Sequencing, One Molecule at a Time.

Brendan M Floyd1, Edward M Marcotte1

  • 1Department of Molecular Biosciences, Center for Systems and Synthetic Biology, University of Texas, Austin, Texas, USA; email: bmfloyd@utexas.edu, marcotte@utexas.edu.

Annual Review of Biophysics
|January 5, 2022
PubMed
Summary
This summary is machine-generated.

Advancing protein characterization, single-molecule protein sequencing (SMPS) methods are emerging to match nucleic acid sequencing

Keywords:
DNA nanotechnologySMPSfluorescencefluorosequencingnanoporesproteomicssingle-molecule protein sequencing

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Last Updated: Oct 7, 2025

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Area of Science:

  • Proteomics
  • Biotechnology
  • Molecular Biology

Background:

  • Protein characterization methods lag behind nucleic acid sequencing in sensitivity, dynamic range, and throughput.
  • Current limitations hinder sensitive medical diagnostics, deep protein quantification, and large-scale measurements.
  • There is a critical need for advanced protein analysis to address biological challenges.

Purpose of the Study:

  • To review and categorize emerging single-molecule protein sequencing (SMPS) technologies.
  • To highlight the potential of SMPS to revolutionize proteomics and related fields.
  • To provide an overview of the current landscape and future directions in SMPS.

Main Methods:

  • Categorization of SMPS technologies into three main approaches: sequencing by degradation, sequencing by transit, and sequencing by affinity.
  • Discussion of diverse experimental approaches, from established to speculative.
  • Review of technologies inspired by high-throughput nucleic acid sequencing.

Main Results:

  • Identification of three primary categories for SMPS: degradation, transit, and affinity-based sequencing.
  • Overview of the spectrum of experimental support for these nascent technologies.
  • Highlighting the potential for significant advancements in proteomics.

Conclusions:

  • SMPS technologies hold promise for overcoming current limitations in protein characterization.
  • These advancements could lead to more sensitive diagnostics and deeper biological insights.
  • The field of proteomics is poised for transformation through the development of SMPS.