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 36
... tube . 5. Incubate at 50-60 ° C for 1 hr . 6. Incubate at 95 ° C for 10 min to inactivate the proteinase . 7. Store frozen . To effect PCR on the lysate produced in step 6 , Protocol A , note that 25 μl of this lysate is about ...
... tube . 5. Incubate at 50-60 ° C for 1 hr . 6. Incubate at 95 ° C for 10 min to inactivate the proteinase . 7. Store frozen . To effect PCR on the lysate produced in step 6 , Protocol A , note that 25 μl of this lysate is about ...
Page 37
... tube and proceed as in step 2 of Protocol B above . The DNA concentration in the final resuspension depends on the number of cells present . 5. If no red blood cells are present , resuspend pellet in PCR buffer with nonionic detergents ...
... tube and proceed as in step 2 of Protocol B above . The DNA concentration in the final resuspension depends on the number of cells present . 5. If no red blood cells are present , resuspend pellet in PCR buffer with nonionic detergents ...
Page 60
... tube to 70 ° C for 3 min for ssDNA templates or for 5 min at 95 ° C for dsDNA templates , then put the tube at 42 ° C for 10 min . If the reaction is performed in microtiter dishes , it may be overlayed by mineral oil to prevent ...
... tube to 70 ° C for 3 min for ssDNA templates or for 5 min at 95 ° C for dsDNA templates , then put the tube at 42 ° C for 10 min . If the reaction is performed in microtiter dishes , it may be overlayed by mineral oil to prevent ...
Contents
PART ONE BASIC METHODOLOGY | 1 |
PCR Amplification of Specific Sequences from | 9 |
Taq DNA Polymerase | 17 |
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 B-globin cDNA cells Cetus chromosome cloned concentration containing cycles deletion denaturant denaturing gradient gel detection DGGE diagnosis direct sequencing disease DNA fragments DNA sequences dNTP dot-blot dystrophin Erlich exon Faloona Figure flanking GC-clamp Gelfand gene 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 sequence temperature template Tris.HCl tube