Lecture

Single Molecule Sequencing: Strategies and Technologies

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In course

MICRO-560: BioMEMS

This course covers the main applications of micro devices for life science and biomedical applications. The course is organized by application topic. It is also covering the basic physical, biological, chemical, technological concepts, which are presented ...

Description

This lecture covers the sequencing strategies and technologies used in single molecule sequencing, including Helicos' sequencing by synthesis method and Pacific Biosciences' real-time sequencing based on zero mode waveguides. It explains the process of adding fluorescent nucleotides one-by-one, cutting away the fluorescent molecule, and repeating the process until the DNA fragments are fully sequenced. The lecture also delves into the use of ZMWs, the incorporation of nucleotides by polymerase, and the diffusion of dye molecules. Additionally, it explores DNA translocation through nanopores, measuring current fluctuation during translocation, and the techniques of 'strand sequencing' and 'exonuclease sequencing' for real-time DNA sequencing.

Instructor

Philippe Renaud

Philippe Renaud is Professor at the Microsystem Laboratory (LMIS4) at EPFL. He is also the scientific director of the EPFL Center of MicroNanoTechnology (CMI). His main research area is related to micronano technologies in biomedical applications (BioMEMS) with emphasis on cell-chips, nanofluidics and bioelectronics. Ph. Renaud is invloved in many scientifics papers in his research area. He received his diploma in physics from the University of Neuchâtel (1983) and his Ph.D. degree from the University of Lausanne (1988). He was postdoctoral fellow at University of California, Berkeley (1988-89) and then at the IBM Zürich Research Laboratory in Switzerland (1990-91). In 1992, he joined the Sensors and Actuators group of the Swiss Center for Electronics and Microtechnology (CSEM) at Neuchâtel, Switzerland. He was appointed assistant professor at EPFL in 1994 and full professor in 1997. In summer 1996, he was visiting professor at the Tohoku University, Japan. Ph. Renaud is active in several scientific committee (scientific journals, international conferences, scientific advisory boards of companies, PhD thesis committee). He is also co-founder of the Nanotech-Montreux conference. Ph. Renaud is committed to valorization of basic research through his involvement in several high-tech start-up companies.

Official source

Ontological neighbourhood

Biology

Genetics: Genomics

Related concepts (30)

DNA sequencing

DNA sequencing is the process of determining the nucleic acid sequence – the order of nucleotides in DNA. It includes any method or technology that is used to determine the order of the four bases: adenine, guanine, cytosine, and thymine. The advent of rapid DNA sequencing methods has greatly accelerated biological and medical research and discovery. Knowledge of DNA sequences has become indispensable for basic biological research, DNA Genographic Projects and in numerous applied fields such as medical diagnosis, biotechnology, forensic biology, virology and biological systematics.

Massive parallel sequencing

Massive parallel sequencing or massively parallel sequencing is any of several high-throughput approaches to DNA sequencing using the concept of massively parallel processing; it is also called next-generation sequencing (NGS) or second-generation sequencing. Some of these technologies emerged between 1993 and 1998 and have been commercially available since 2005. These technologies use miniaturized and parallelized platforms for sequencing of 1 million to 43 billion short reads (50 to 400 bases each) per instrument run.

Third-generation sequencing

Third-generation sequencing (also known as long-read sequencing) is a class of DNA sequencing methods currently under active development. Third generation sequencing technologies have the capability to produce substantially longer reads than second generation sequencing, also known as next-generation sequencing. Such an advantage has critical implications for both genome science and the study of biology in general. However, third generation sequencing data have much higher error rates than previous technologies, which can complicate downstream genome assembly and analysis of the resulting data.

Shotgun sequencing

In genetics, shotgun sequencing is a method used for sequencing random DNA strands. It is named by analogy with the rapidly expanding, quasi-random shot grouping of a shotgun. The chain-termination method of DNA sequencing ("Sanger sequencing") can only be used for short DNA strands of 100 to 1000 base pairs. Due to this size limit, longer sequences are subdivided into smaller fragments that can be sequenced separately, and these sequences are assembled to give the overall sequence.

Exome sequencing

Exome sequencing, also known as whole exome sequencing (WES), is a genomic technique for sequencing all of the protein-coding regions of genes in a genome (known as the exome). It consists of two steps: the first step is to select only the subset of DNA that encodes proteins. These regions are known as exons—humans have about 180,000 exons, constituting about 1% of the human genome, or approximately 30 million base pairs. The second step is to sequence the exonic DNA using any high-throughput DNA sequencing technology.

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