PCR Protocols: A Guide to Methods and ApplicationsMichael A. Innis The broad utility of the polymerase chain reaction (PCR) method is now within the reach of every researcher. Designed for use at the laboratory bench, this is the most comprehensive manual on PCR available today. Over 50 chapters provide precise instructions on procedures, with advice on primer design. All of the techniques described, from amplification and direct sequencing of genomic DNA through cDNA cloning and quantitation of mRNA, are tested, current, and supplemented with helpful notes and illustrations. You'll also learn how to: * Optimize novel applications * Avoid many cumbersome molecular biological techniques * Set up the laboratory to avoid contamination |
From inside the book
Results 1-3 of 69
Page 5
... dNTP is recommended . The stability of the dNTPs during repeated cycles of PCR is such that approximately 50 % remains as dNTP after 50 cycles ( Corey Levenson , personal communication ) . Deoxynucleotide concentrations between 20 and ...
... dNTP is recommended . The stability of the dNTPs during repeated cycles of PCR is such that approximately 50 % remains as dNTP after 50 cycles ( Corey Levenson , personal communication ) . Deoxynucleotide concentrations between 20 and ...
Page 6
... dNTP con- centration appropriate for the length and composition of the target sequence ; e.g. , 20 μM each dNTP in a 100 - μl reaction is theoreti- cally sufficient to synthesize 2.6 μg of DNA or 10 pmol of a 400 - bp sequence ...
... dNTP con- centration appropriate for the length and composition of the target sequence ; e.g. , 20 μM each dNTP in a 100 - μl reaction is theoreti- cally sufficient to synthesize 2.6 μg of DNA or 10 pmol of a 400 - bp sequence ...
Page 136
... dNTP concentration . In addition , the synthesis rate of Taq poly- merase decreases by as much as 20 to 30 % as the total dNTP con- centration is increased to 4 to 6 mM , despite optimization of magne- sium concentration for each dNTP ...
... dNTP concentration . In addition , the synthesis rate of Taq poly- merase decreases by as much as 20 to 30 % as the total dNTP con- centration is increased to 4 to 6 mM , despite optimization of magne- sium concentration for each dNTP ...
Contents
Amplification of Genomic DNA | 13 |
Amplification of RNA | 21 |
Rapid Amplification of cDNA Ends | 28 |
Copyright | |
34 other sections not shown
Other editions - View all
Common terms and phrases
1990 by Academic Acad Academic Press agarose gel aliquots alleles amplification products amplified DNA analysis annealing Applications Copyright assay cDNA cells chromosome cloning concentration containing cycles denaturation detection digestion DNA fragments DNA sequences dNTP dot blot EDTA electrophoresis enzymatic amplification ethanol ethidium bromide extraction Faloona form reserved gene genetic genomic genomic DNA Guide to Methods H. A. Erlich Higuchi hybridization Incubate labeled lanes Literature Cited membrane Methods and Applications MgCl2 minutes molecular molecules mRNA mutations Natl Nucleic Acids Res nucleotide oligomer oligonucleotide probes PCR amplification PCR primers PCR product PCR Protocols Perkin-Elmer Cetus phage pmol polymerase chain reaction polymorphisms primer primer pair procedure purified quences reagents recombinant region restriction enzymes reverse transcriptase rights of reproduction rRNA Saiki Scharf single-stranded Sninsky solution Southern blot specific strand synthesis Taq DNA polymerase Taq polymerase temperature template tion tissue transcription Tris-HCl pH tube virus vitro