PCR Technology: Principles and Applications for DNA AmplificationPolymerase chain reaction (PCR) technology is a revolutionary innovation which enables scientists to rapidly generate large amounts of genetic material from a slight trace which would otherwise be too small to analyze. With applications in both research and diagnostics, PCR is becoming a standard procedure in biotechnology and medical diagnostic laboratories. This book is an introduction and guide to the new technology, covering the basic methodologies and their applications in research and medicine, emphasizing practical aspects. Each chapter is written by pioneers in the field and most include detailed protocols and favorite PCR "recipes". Students and researchers in all areas of biotechnology and molecular biology will find this book the introduction to PCR they've been looking for. |
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Page 36
3 . Resuspend cells in 10 ml PBS and repeat centrifugation / wash 4 . Resuspend
in a volume of " PCR buffer w / nonionic detergents and Proteinase K " to give
about 6x10 cells per ml . Transfer to 1 . 5 - ml Eppendorf microcentrifuge tube . 5 .
3 . Resuspend cells in 10 ml PBS and repeat centrifugation / wash 4 . Resuspend
in a volume of " PCR buffer w / nonionic detergents and Proteinase K " to give
about 6x10 cells per ml . Transfer to 1 . 5 - ml Eppendorf microcentrifuge tube . 5 .
Page 58
The reaction is performed in 10 ul sequencing buffer ( 50 mM KCI , 50 mM Tris .
HCI , pH 8 . 0 , 5 mM MgCl2 , 10 mm dithiothreitol ) . Heat the reaction on ice for
10 min and then transfer the tube to ice . Transfer 2 . 2 ul of the mixture to each of
...
The reaction is performed in 10 ul sequencing buffer ( 50 mM KCI , 50 mM Tris .
HCI , pH 8 . 0 , 5 mM MgCl2 , 10 mm dithiothreitol ) . Heat the reaction on ice for
10 min and then transfer the tube to ice . Transfer 2 . 2 ul of the mixture to each of
...
Page 221
Digestion Buffer : 10 mM Tris . HCI , pH 8 . 0 , 10 mM EDTA , 50 mM NaCl , 2 %
SDS Water - saturated n - butanol • II . Digestion Procedures A . Blood Digestion
Suspend buffy coat , 0 . 1 ml whole blood , or blood stain in 0 . 5 ml digestion ...
Digestion Buffer : 10 mM Tris . HCI , pH 8 . 0 , 10 mM EDTA , 50 mM NaCl , 2 %
SDS Water - saturated n - butanol • II . Digestion Procedures A . Blood Digestion
Suspend buffy coat , 0 . 1 ml whole blood , or blood stain in 0 . 5 ml digestion ...
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Contents
PART ONE BASIC METHODOLOGY | 1 |
The Design and Optimization of the PCR | 7 |
Taq DNA Polymerase | 17 |
Copyright | |
14 other sections not shown
Other editions - View all
PCR Technology: Principles and Applications for DNA Amplification Henry Erlich No preview available - 1989 |
Common terms and phrases
Acad Acids activity addition alleles allow Alu repeats amount amplification analysis annealing applications approach assay base blood buffer carried cDNA cell changes Chapter clones concentration containing cycles deletion denaturation described detection determined diagnosis digestion direct discussed disease DNA fragments DNA polymerase DNA sequence dNTP domain effect efficient electrophoresis enzyme Erlich example exon extension Figure gene genetic genomic human hybridization identified increase individual initial isolation known lane limited loci locus markers melting method molecular Mullis mutations Nature observed obtained oligonucleotide PCR product performed polymorphic positive possible preparation present primers probes problem Proc procedure Protocol reaction recombination region restriction Saiki samples Science separate shown single specific sperm step strand studies Taq polymerase techniques temperature template tube usually yield
References to this book
Bibliographic information
| Title | PCR Technology: Principles and Applications for DNA Amplification Breakthroughs in Molecular Biology Series |
| Author | Henry A. Erlich |
| Editor | Henry A. Erlich |
| Edition | illustrated, reprint |
| Publisher | Oxford University Press, 1992 |
| Original from | the University of Michigan |
| Digitized | 28 Jul 2008 |
| ISBN | 0195098757, 9780195098754 |
| Length | 246 pages |
| Subjects | › › Literary Criticism / European / English, Irish, Scottish, Welsh |
| Export Citation | BiBTeX EndNote RefMan |