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Related Concept Videos

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.
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The...
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.
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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. 
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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...

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Related Experiment Video

Updated: Jun 27, 2026

High-throughput Identification of Gene Regulatory Sequences Using Next-generation Sequencing of Circular Chromosome Conformation Capture (4C-seq)
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Sequencing by Cyclic Ligation and Cleavage (CycLiC) directly on a microarray captured template.

Kalim U Mir1, Hong Qi, Oleg Salata

  • 1The Wellcome Trust Centre for Human Genetics, Oxford University, Oxford, UK. kalim.mir@well.ox.ac.uk

Nucleic Acids Research
|November 19, 2008
PubMed
Summary
This summary is machine-generated.

A new massively scalable biochemistry, Cyclical Ligation and Cleavage (CycLiC), enables contiguous base sequencing. This method, applied to microarray-captured templates, accurately reads DNA sequences for genomic applications.

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Area of Science:

  • Genomics and Molecular Biology
  • Next-Generation Sequencing Technologies

Background:

  • Current next-generation sequencing (NGS) methods require enhancement for both whole-genome and targeted resequencing applications.
  • A need exists for scalable, efficient, and cost-effective sequencing platforms applicable to diverse genomic studies.

Purpose of the Study:

  • To introduce and validate a novel massively scalable biochemistry for contiguous base sequencing, termed Cyclical Ligation and Cleavage (CycLiC).
  • To demonstrate the direct application of CycLiC to DNA templates immobilized on a microarray platform.
  • To assess the feasibility of CycLiC for accurate DNA sequence determination in a high-throughput format.

Main Methods:

  • Development of Cyclical Ligation and Cleavage (CycLiC) biochemistry utilizing four color-coded DNA/RNA chimeric oligonucleotide libraries (OL).
  • Step-wise primer extension along a DNA template, with each cycle involving OL ligation, label detection for base identification, and cleavage of the label and undetermined bases.
  • Immobilization of DNA templates onto a microarray via hybridization and subsequent sequencing using the CycLiC method.

Main Results:

  • Proof-of-principle demonstration confirming that the CycLiC method functions as designed.
  • Accurate contiguous base sequencing of DNA templates captured on a microarray probe was achieved.
  • The CycLiC method proved amenable to massive scale-up, miniaturization, and automation.

Conclusions:

  • The CycLiC method provides a scalable and accurate approach for contiguous base sequencing.
  • Microarray implementation of CycLiC enables simultaneous selection and sequencing of numerous genomic regions on a single platform.
  • The use of common reagents facilitates further development and adoption by the research community.