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 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 187
... Incubate 37 ° C for 1 hr , then add 30 μl of 0.7M NaOH , 40 mM EDTA , mix gently , and incubate at 65 ° C for 10 min . 4. Add 5 μl of 2 M ammonium ... incubation is extended for 7 min . 5. Analyze double - stranded PCR product by agarose gel.
... Incubate 37 ° C for 1 hr , then add 30 μl of 0.7M NaOH , 40 mM EDTA , mix gently , and incubate at 65 ° C for 10 min . 4. Add 5 μl of 2 M ammonium ... incubation is extended for 7 min . 5. Analyze double - stranded PCR product by agarose gel.
Page 221
... Incubate at least 1 h at 56 ° C . Remove blood stain substrate . Add 0.5 ml phenol / chloroform / isoamyl alcohol . Vortex . Spin in microcentrifuge 2 min . Transfer top ( aqueous ) phase to new tube . Repeat extraction until interface ...
... Incubate at least 1 h at 56 ° C . Remove blood stain substrate . Add 0.5 ml phenol / chloroform / isoamyl alcohol . Vortex . Spin in microcentrifuge 2 min . Transfer top ( aqueous ) phase to new tube . Repeat extraction until interface ...
Contents
PART ONE BASIC METHODOLOGY | 1 |
The Design and Optimization of the PCR | 7 |
PCR Amplification of Specific Sequences from | 9 |
Copyright | |
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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 disease DNA fragments DNA sequence dNTP dot-blot dystrophin Erlich exon Faloona Figure flanking GC-clamp Gelfand genetic genomic genomic DNA heteroduplexes Higuchi HPRT human hybridization incubation individual 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