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By Ehrenborg R.

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TaqMan is used for mutation detection and for screening specific subtypes of microbial pathogens. , in the cytochrome P450 genes) (11). It is anticipated that this will lead to the identification of novel drug targets and aid in the production of individually tailored “designer” drugs. 6. Quantitative and Real-Time PCR Real-Time PCR requires the use of special DNA cyclers such as the Roche Lightcycler that couples PCR with fluorescent detection, thereby enabling the detection of PCR product as it is synthesized.

Humana, Totowa, NJ 3. Fischer SG, Lerman LS (1983) DNA fragments differing by a single base-pair substitution are separated in denaturing gradient gels: correspondence with melting theory. Proc Natl Acad Sci USA 80:1579–1583 4. Huber CG, Oefner PJ, Bonn GK (1995) Rapid and accurate sizing of DNA fragments by ion-pair chromatography on alkylated nonporous poly(styrenedivinylben zene) particles. Anal Chem 67:578–585 5. Cotton RGH, Rodrigues NR, Campbell RD (1988) Reactivity of cytosine and thymine in single base-pair mismatches with hydroxylamine and osmium tetroxide Chapter 3 Principles and Medical Applications of the Polymerase Chain Reaction 6.

Large exons are usually amplified as several overlapping segments. 1. Reverse Transcriptase PCR A number of screening methods are able to analyze 1 kb or more at a time, in which case, PCR-amplified complementary or cDNA may be used. This is DNA synthesized using messenger RNA (mRNA) as a template mediated by the enzyme reverse transcriptase (see Fig. 4). This procedure, referred to as RT-PCR, enables the screening of multiple contiguous exons in a single analysis, which enables the identification of gene rearrangements and splicing Chapter 3 Principles and Medical Applications of the Polymerase Chain Reaction Fig.

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A Bijective Answer to a Question of Zvonkin by Ehrenborg R.

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