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
... dNTP ) , but in some circumstances , different amounts of Mg2 + may prove to be necessary ( Figure 2 ) . Generally , excess Mg2 + will result in the accumulation of non - specific amplification products and insufficient Mg2 + will ...
... dNTP ) , but in some circumstances , different amounts of Mg2 + may prove to be necessary ( Figure 2 ) . Generally , excess Mg2 + will result in the accumulation of non - specific amplification products and insufficient Mg2 + will ...
Page 18
... dNTP concentration . In addition , at the correspondingly optimal magnesium concentration , the synthesis rate of Taq polymerase decreases by as much as 20-30 % as the total dNTP concentration is increased to 4-6 mM . This observation ...
... dNTP concentration . In addition , at the correspondingly optimal magnesium concentration , the synthesis rate of Taq polymerase decreases by as much as 20-30 % as the total dNTP concentration is increased to 4-6 mM . This observation ...
Page 59
... dNTP except dATP ) . Add 0.5 μl of a❝S - thio dATP ( 1000 Ci / mmol ) , 10 μCi / ul . Add 2.0 μl of T7 DNA polymerase ( diluted 1/8 in TE ) . · The mixture is left at room temperature for 5 min . Add 3.5 μl of the labeling reaction to ...
... dNTP except dATP ) . Add 0.5 μl of a❝S - thio dATP ( 1000 Ci / mmol ) , 10 μCi / ul . Add 2.0 μl of T7 DNA polymerase ( diluted 1/8 in TE ) . · The mixture is left at room temperature for 5 min . Add 3.5 μl of the labeling reaction to ...
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