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Related Concept Videos

Sanger Sequencing01:57

Sanger Sequencing

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...
Next-generation Sequencing03:00

Next-generation Sequencing

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.
Maxam-Gilbert Sequencing01:05

Maxam-Gilbert Sequencing

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...
RNA-seq03:21

RNA-seq

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 microarray-based...

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Related Experiment Video

Updated: Jun 8, 2026

An Ultrahigh-throughput Microfluidic Platform for Single-cell Genome Sequencing
10:00

An Ultrahigh-throughput Microfluidic Platform for Single-cell Genome Sequencing

Published on: May 23, 2018

Single molecule sequencing with a HeliScope genetic analysis system.

John F Thompson1, Kathleen E Steinmann

  • 1Helicos BioSciences, Cambridge, Massachusetts, USA.

Current Protocols in Molecular Biology
|October 5, 2010
PubMed
Summary
This summary is machine-generated.

Helicos Single Molecule Sequencing (SMS) offers unbiased genome analysis by directly sequencing cellular nucleic acids. This method avoids biases from ligation or PCR, enabling accurate quantitation and sequence information with minimal sample input.

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Area of Science:

  • Genomics
  • Molecular Biology
  • Biotechnology

Background:

  • Traditional sequencing methods often introduce biases through processes like ligation and PCR amplification.
  • These biases can affect quantitative accuracy and representation of genomic regions, particularly those with extreme GC content or varying sizes.

Purpose of the Study:

  • To describe the principles and methods of the Helicos™ Single Molecule Sequencing (SMS) system.
  • To highlight the advantages of direct nucleic acid sequencing for unbiased genomic analysis and gene expression profiling.

Main Methods:

  • Direct sequencing of cellular nucleic acids without ligation or PCR amplification.
  • DNA preparation involves shearing, poly(A) tailing, and hybridization to an oligo(dT)-coated flow cell.
  • Sequencing-by-synthesis of billions of molecules in parallel.
  • Gene expression analysis via first-strand cDNA (RNA-Seq) or direct RNA hybridization and sequencing.

Main Results:

  • Helicos SMS provides accurate quantitation and sequence information.
  • The method avoids GC-content and size biases inherent in other technologies.
  • Requires significantly less starting material compared to alternative methods.
  • Enables direct RNA sequencing for highly accurate gene expression measurements.

Conclusions:

  • Helicos SMS offers a powerful, unbiased approach to genome and transcriptome analysis.
  • The technology simplifies sample preparation and reduces bias, leading to more accurate biological insights.
  • Direct RNA sequencing represents a significant advancement for precise gene expression quantification.