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 8
... base distribution and with a GC content similar to that of the fragment being amplified . Try to avoid primers with stretches of polypurines , polypyrimidines , or other unusual sequences . Figure 1. Amplification of B - globin ...
... base distribution and with a GC content similar to that of the fragment being amplified . Try to avoid primers with stretches of polypurines , polypyrimidines , or other unusual sequences . Figure 1. Amplification of B - globin ...
Page 10
... bases in length and the optimal amount to use in an amplification will vary . Longer primers may be synthesized but are seldom necessary . Sequences not complementary to the template can be added to the 5 ' - end of the primers . These ...
... bases in length and the optimal amount to use in an amplification will vary . Longer primers may be synthesized but are seldom necessary . Sequences not complementary to the template can be added to the 5 ' - end of the primers . These ...
Page 13
... bases or less . The polymerase retains significant activity at lower temperatures ' and complete extension will occur during the thermal transition from annealing to denaturation . ) The ramp time , or time taken to change from one ...
... bases or less . The polymerase retains significant activity at lower temperatures ' and complete extension will occur during the thermal transition from annealing to denaturation . ) The ramp time , or time taken to change from one ...
Page 14
... base oligonucleotide primers with about 50 % GC content ; even higher temperatures may be necessary to increase ... bases are available and an annealing temperature around 40-45 ° C is needed . However , primers of that length are ...
... base oligonucleotide primers with about 50 % GC content ; even higher temperatures may be necessary to increase ... bases are available and an annealing temperature around 40-45 ° C is needed . However , primers of that length are ...
Page 54
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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