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
... PCR reaction . It is very important that the reaction reaches a temperature at which complete strand separation occurs . A temperature of about 94 ° C should be adequate in most cases . As soon as the sample reaches 94 ° C , it can be ...
... PCR reaction . It is very important that the reaction reaches a temperature at which complete strand separation occurs . A temperature of about 94 ° C should be adequate in most cases . As soon as the sample reaches 94 ° C , it can be ...
Page 58
... reaction for 15 min . Dry down the reaction , resuspend it in 3 μl water and add 3 μl formamide - dye stop solution ( 90 % formamide , 20 mM EDTA , pH 8.0 and 0.05 % each of the dyes xylene cyanol and bromophenol blue ) . 2. Kinased ...
... reaction for 15 min . Dry down the reaction , resuspend it in 3 μl water and add 3 μl formamide - dye stop solution ( 90 % formamide , 20 mM EDTA , pH 8.0 and 0.05 % each of the dyes xylene cyanol and bromophenol blue ) . 2. Kinased ...
Page 93
... PCR reaction . It may be useful to titrate the primer oligonucleotides in the PCR reaction to find the lowest amount that can be used to give a " good " amplified product . We have found that a large excess of primers in the reaction ...
... PCR reaction . It may be useful to titrate the primer oligonucleotides in the PCR reaction to find the lowest amount that can be used to give a " good " amplified product . We have found that a large excess of primers in the reaction ...
Contents
PART ONE BASIC METHODOLOGY | 1 |
The Design and Optimization of the PCR | 7 |
PCR Amplification of Specific Sequences from | 9 |
Copyright | |
17 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 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