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Since the independent invention of DNA sequencing by Sanger and by
Gilbert 30 years ago, it has grown from a small scale technique
capable of reading several kilobase-pair of sequence per day into
today's multibillion dollar industry. This growth has spurred the
development of new sequencing technologies that do not involve
either electrophoresis or Sanger sequencing chemistries. Sequencing
by Synthesis (SBS) involves multiple parallel micro-sequencing
addition events occurring on a surface, where data from each round
is detected by imaging.
New High Throughput Technologies for DNA Sequencing and Genomics is
the second volume in the Perspectives in Bioanalysis series, which
looks at the electroanalytical chemistry of nucleic acids and
proteins, development of electrochemical sensors and their
application in biomedicine and in the new fields of genomics and
proteomics. The authors have expertly formatted the information for
a wide variety of readers, including new developments that will
inspire students and young scientists to create new tools for
science and medicine in the 21st century.
Reviews of complementary developments in Sanger and SBS sequencing
chemistries, capillary electrophoresis and microdevice integration,
MS sequencing and applications set the framework for the book.
* 'Hot Topic' with DNA sequencing continuing as a major research
activity in many areas of life science and medicine.
* Bringing together new developments in DNA sequencing
technology
* Reviewing issues relevant to the new applications used
An outstanding panel of hands-on experts and developers of CE
equipment describe in step-by-step fashion their best cutting-edge
methods for the detection and analysis of DNA mutations and
modifications, ranging from precise DNA loci to entire genomes of
organisms. This first volume of the set, Introduction to the
Capillary Electrophoresis of Nucleic Acids, covers the practical
and theoretical considerations behind the use of capillary
electrophoresis for the analysis of small oligonucleotides and
modified nucleotides. Along with detailed instructions ensuring
ready reproducibility, these protocols offer time-tested advice on
instrumentation, signal detection, the capillary environment, and
the integration of mass spectrometry with CE. Several chapters are
devoted to the analysis of small therapeutic oligonucleotides,
nucleosides, and ribonucleotides by CE. The companion volume,
Practical Applications of Capillary Electrophoresis, addresses
techniques for high-throughput analysis of DNA fragments using SNP
detection, mutation detection, DNA sequencing methods, and
DNA-ligand interactions. Comprehensive and up-to-date, the paired
volumes of Capillary Electrophoresis of Nucleic Acids offer an
authoritative guide with easy access to fast, versatile, reliable,
and powerful technologies for all those basic and clinical
investigators analyzing DNA variation today.
The development of PCR, which enables extremely small amounts of
DNA to be amplified, led to the rapid development of a multiplicity
of a- lytical procedures to utilize this new resource for analysis
of genetic variation and for the detection of disease causing
mutations. The advent of capillary electrophoresis (CE), with its
power to separate and analyze very small amounts of DNA, has also
stimulated researchers to develop analytical procedures for the CE
format. The advantages of CE in terms of speed and reproducibility
of analysis are manifold. Further, the high sensitivity of
detection, and the ab- ity to increase sample throughput with
parallel analysis, has led to the creation of a full range of
analysis of DNA molecules, from modified DNA-adducts and
single-strand oligonucleotides through to PCR-amplified DNA
fragments and whole chromosomes. Capillary Electrophoresis of
Nucleic Acids focuses on such analytical protocols, which can be
used for detection and analysis of mutations and modification, from
precise DNA loci through to entire genomes of organisms. Important
practical considerations for CE, such as the choice of separation
media, electrophoresis conditions, and the influence of buffer
additives and dyes on DNA mobility, are discussed in several key
chapters and within particular applications.
The development of PCR, which enables extremely small amounts of
DNA to be amplified, led to the rapid development of a multiplicity
of a- lytical procedures that permit use of this new resource for
the analysis of genetic variation and for the detection of
disease-causing mutations. The advent of capillary electrophoresis
(CE), with its power to separate and a- lyze very small amounts of
DNA, has also stimulated researchers to develop analytical
procedures for the CE format. The advantages of CE in terms of
speed and reproducibility of analyses are manifold. Furthermore,
the high s- sitivity of detection, and the ability to increase
sample throughput with par- lel analysis, has led to the creation
of a full range of analysis of DNA molecules, from modified DNA
adducts and single-strand oligonucleotides through PCR-amplified
DNA fragments and whole chromosomes. Capillary Elect- phoresis of
Nucleic Acids focuses on analytical protocols that can be used for
detection and analysis of mutations and modification, from precise
DNA loci through entire genomes of organisms. Important practical
considerations for CE, such as the choice of separation media,
electrophoresis conditions, and the influence of buffer additives
and dyes on DNA mobility, are discussed in several key chapters and
within particular applications.
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