LEADER 05527nam 2200733 a 450 001 9910455737203321 005 20200520144314.0 010 $a0-19-156569-5 010 $a1-280-37548-5 010 $a9786610375486 010 $a0-585-48413-9 035 $a(CKB)111087026789960 035 $a(EBL)1044783 035 $a(OCoLC)814379107 035 $a(SSID)ssj0000207294 035 $a(PQKBManifestationID)12030395 035 $a(PQKBTitleCode)TC0000207294 035 $a(PQKBWorkID)10237156 035 $a(PQKB)11380054 035 $a(MiAaPQ)EBC1044783 035 $a(Au-PeEL)EBL1044783 035 $a(CaPaEBR)ebr10612572 035 $a(CaONFJC)MIL37548 035 $a(EXLCZ)99111087026789960 100 $a19971204d1998 uy 0 101 0 $aeng 135 $aur|n|---||||| 181 $ctxt 182 $cc 183 $acr 200 00$aMutation detection$b[electronic resource] $ea practical approach /$fedited by R.G.H. Cotton, E. Edkins, and S. Forrest 210 $aOxford ;$aNew York $cIRL Press at Oxford University Press$d1998 215 $a1 online resource (263 p.) 225 0$aPractical approach series ;$v188 300 $aDescription based upon print version of record. 311 $a0-19-963657-5 311 $a0-19-963656-7 320 $aIncludes bibliographical references and index. 327 $aCover; Contents; List of Contributors; Abbreviations; Introduction; References; 1. Single-strand conformation polymorphism analysis; 1. Introduction; 2. PCR-SSCP using polyacrylamide slab gel; PCR Optimization and primer design; Pre-amplification and isolation by agarose gel electrophoresis; PCR using [[Sup(32)]P]deoxynucleotide triphosphate; Removal of 3' appendage; SSCP gel electrophoresis; Interpretation of autoradiogram; Re-amplification and direct sequencing; Gel matrices other than polyacrylamide; Restriction endonuclease fingerprinting and dideoxy fingerprinting 327 $a3. Fluorescent SSCP in an automated DNA sequencerPrimer design in post-PCR fluorescent labelling; Fluorescent labelling by 3' exchange reaction; SSCP in capillary electrophoresis (CE-SSCP); Data processing; Acknowledgements; References; 2. Single-stranded conformation polymorphism and heteroduplex analysis; 1. Introduction; 2. Optimization of the PCR reaction; 3. SSCP sample prepration; 4. Optimization of SSCP/HA detection; 5. Multiplexing; 6. Interpretation of results; 7. Applications; 8. Other methods; References 327 $a3. Comprehensive mutation detection with denaturing gradient gel electrophoresis1. Introduction; The scope of DGGE, its distinctive capabilities, and the nature of results; 2. Background; 3. Basic principle, the physical properties of DNA; 4. Overview of the procedures in searching for mutants; Defining segments for scrutiny; Sample preparation; Gradient and velocity separations; Features of the gel patterns; Discrimination of zygozygosity; Comments; 5. Use of the psoralen cross-link as a clamp; The psoralen protocol; 6. Computational tools; What is a meltmap?; Meltmap protocol 327 $aPredicting electrophoretic separationsComputer operations for MUTRAV; 7. Other members of the DGGE family; Gel separations in a uniform, partially denaturing environment; Capillary electrophoresis; The thermal gradient; The temperature ramp; 2D length and gradient separations; 8. End notes; Acknowledgments; References; 4. Cleavage using RNase to detect mutations; 1. Introduction; 2. RNase protection assay for mutation detection; Evaluation of the sensitivity; Source material; PCR for RNase protection assay; RNA probe preparation; RNase protection; Detection of digested probe 327 $aMutation detection by sequencing of the PCR productsOther modified methodologies for mutation detection; Acknowledgements; References; 5. Cleavage of mismatched bases using chemical reagents; 1. Introduction; 2. Basic procedures; Comments on the basic procedures; 3. Ultra fast chemical mismatch detection; Labelling; Solid phase; Comments; References; 6. Mutation detection using T4 endonuclease VII; 1. Introduction; 2. The biology of Endo VII; The role of Endo VII in vivo; Characterization of Endo VII; Action of Endo VII on heteroduplex DNA; 3. Use of Endo VII for mutation detection 327 $aEnzyme mismatch cleavage 330 $aMutation detection is increasingly undertaken in a wide spectrum of research areas: in medicine it is fundamental in isolating disease genes and diagnbosis, and is especially important in cancer research; in biology, commercially important genes can be identified by the mutations they contain. But mutation detection is time-consuming and expensive. This volume offers the latest tried and tested protocols for a range of detection methods, from the labs of the leading researchers inthe field. 410 0$aPractical Approach Series 606 $aMutation (Biology)$vLaboratory manuals 606 $aMolecular genetics$vLaboratory manuals 606 $aChromosome abnormalities$vLaboratory manuals 608 $aElectronic books. 615 0$aMutation (Biology) 615 0$aMolecular genetics 615 0$aChromosome abnormalities 676 $a576.5/49 701 $aCotton$b Richard G. H$0945901 701 $aEdkins$b E$g(Edward)$0945902 701 $aForrest$b S$g(Sue)$0945903 801 0$bMiAaPQ 801 1$bMiAaPQ 801 2$bMiAaPQ 906 $aBOOK 912 $a9910455737203321 996 $aMutation detection$92136728 997 $aUNINA