PCR Technology: Principles and Applications for DNA AmplificationThis is an introduction to the methods and applications of polymerase chain reaction (PCR) technology, a technology developed by Erlich's group at Cetus and Cetus, and is expected to be used in all biology laboratories worldwide within the next few years. |
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Page 11
... incubating the samples at three temperatures corresponding to the three steps in a cycle of amplification - denaturation , annealing , and extension . This cycling can be accomplished either manually with pre - set water baths , or ...
... incubating the samples at three temperatures corresponding to the three steps in a cycle of amplification - denaturation , annealing , and extension . This cycling can be accomplished either manually with pre - set water baths , or ...
Page 13
... incubation at 70-75 ° C varies according to the length of target being amplified ; allowing 1 min for each kilobase of sequence is almost certainly excessive , but it is a good place to begin . Shorter times should be tried once the ...
... incubation at 70-75 ° C varies according to the length of target being amplified ; allowing 1 min for each kilobase of sequence is almost certainly excessive , but it is a good place to begin . Shorter times should be tried once the ...
Page 14
... incubation at the annealing temperature is not required . ( The 20 sec denaturation and annealing incubation times used with the Step - Cycle program on the Thermal Cycler is the amount of time it takes for a 100 - μl reaction in a 0.5 ...
... incubation at the annealing temperature is not required . ( The 20 sec denaturation and annealing incubation times used with the Step - Cycle program on the Thermal Cycler is the amount of time it takes for a 100 - μl reaction in a 0.5 ...
Page 15
... incubation time during the annealing and extension steps will limit the opportunities for mispriming and extension by molecules of otherwise idle DNA polymerase . Reducing primer and enzyme concentrations also serves to limit mispriming ...
... incubation time during the annealing and extension steps will limit the opportunities for mispriming and extension by molecules of otherwise idle DNA polymerase . Reducing primer and enzyme concentrations also serves to limit mispriming ...
Page 24
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Contents
1 | |
9 | |
17 | |
PCR Automation | 23 |
Simple and Rapid Preparation | 31 |
PART TWO RESEARCH APPLICATIONS | 39 |
Using PCR to Engineer DNA | 61 |
Mutation Detection by PCR GCClamps | 71 |
The Use of Repeat Sequence | 113 |
A New Approach to Constructing Genetic | 119 |
Evolutionary Analysis via PCR | 137 |
PART THREE MEDICAL APPLICATIONS | 149 |
Diagnosis of New Mutation Diseases Using | 171 |
Applications of PCR to the Analysis | 209 |
Detection of ras Oncogenes Using PCR | 225 |
Application of PCR to the Detection | 235 |
Other editions - View all
PCR Technology: Principles and Applications for DNA Amplification Henry Erlich No preview available - 1989 |
Common terms and phrases
Acad Acids Res agarose alleles amplification amplification products amplified DNA analysis annealing assay automated cDNA cells Cetus chromosome cloned concentration containing cycles deletion denaturant denaturing gradient gel detection DGGE diagnosis direct sequencing DNA fragments DNA sequences dNTP dot-blot dystrophin Erlich exon Faloona Figure flanking GC-clamp gel electrophoresis Gelfand genetic genomic genomic DNA heteroduplexes Higuchi HPRT human hybridization incubation inverse PCR lane Lerman loci locus markers melting behavior melting domain method misincorporation mismatch molecular molecules mRNA Mullis mutations Natl Nucl nucleotide oligonucleotide primers PCR amplification PCR buffer PCR fragments PCR primers PCR product PCR reaction pmol polymerase chain reaction polymorphism primers Proc procedure protein Proteinase Proteinase K Protocol recombination region restriction enzyme reverse transcriptase Saiki Scharf sequencing reactions single-stranded specific ssDNA strands synthesis Taq DNA polymerase Taq polymerase target DNA target sequence temperature template Tris.HCl tube