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Produktbild: Techniques in Animal Cytogenetics

Techniques in Animal Cytogenetics

Aus der Reihe Principles and Practice

Fr. 72.90

inkl. gesetzl. MwSt., Versandkostenfrei

Beschreibung

Produktdetails

Einband

Taschenbuch

Erscheinungsdatum

01.11.2011

Herausgeber

Paul Popescu + weitere

Verlag

Springer Berlin

Seitenzahl

230

Maße (L/B/H)

23.5/15.5/1.5 cm

Gewicht

400 g

Auflage

Softcover reprint of the original 1st ed. 2000

Übersetzt von

R. Popescu

Sprache

Englisch

ISBN

978-3-642-64095-7

Beschreibung

Produktdetails

Einband

Taschenbuch

Erscheinungsdatum

01.11.2011

Herausgeber

Verlag

Springer Berlin

Seitenzahl

230

Maße (L/B/H)

23.5/15.5/1.5 cm

Gewicht

400 g

Auflage

Softcover reprint of the original 1st ed. 2000

Übersetzt von

R. Popescu

Sprache

Englisch

ISBN

978-3-642-64095-7

Herstelleradresse

Springer-Verlag KG
Sachsenplatz 4-6
1201 Wien
AT

Email: GPSR Kontakt

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  • Produktbild: Techniques in Animal Cytogenetics
  • I Preparation of Chromosome Spreads.- I.1 Cell culture techniques.- I.1.1 Lymphocyte culture.- Principle.- Protocol for whole blood.- Protocol for lymphocytes isolated by sedimentation.- Protocol for lymphocyte isolation by density gradient centrifugation.- Remarks.- I.1.2 Fibroblast cultures.- I.1.2.1 Cultures established from tissue fragments.- Principle.- Protocol.- Remarks.- I.1.2.2 Initiation of cell cultures from bird embryos.- Principle.- Protocol.- Remarks.- I.1.2.3 Cell counting.- I.1.2.4 Cell storage.- Principle.- Protocol.- I.2 Preparation of chromosomes in prophase or prometaphase.- I.2.1 Single or double synchronisation using thymidine.- Principle.- Protocol for lymphocytes (single synchronisation).- Protocol for fibroblasts (double synchronisation).- Remarks.- I.2.2 Synchronisation by amethopterin.- Principle.- Protocol.- Remarks.- I.2.3 Synchronisation by 5-fluorodeoxiuridine.- Principle.- I.2.4 Synchronisation by 5-bromodeoxyuridine or BrdU.- Principle.- I.3 Direct techniques and very short term development cultures.- I.3.1 Study of the bone marrow.- Protocol.- Remarks.- I.3.2 Study of bird chromosomes from feather pulp.- Protocol.- Remarks.- I.3.3 Study of fish chromosomes.- Protocol for culture of blood lymphocytes.- Protocol for culture of lymphocytes from kidney or spleen.- Protocol for chromosomal banding by BrdU incorporation in live fish.- Remarks.- I.3.4 Study of insect chromosomes.- I.3.4.1 Preparation of metaphase spreads from embryo cells.- Protocol.- I.3.4.2 Preparation of metaphase spreads from gonads.- Protocol.- I.3.4.3 Chromosome analysis from other tissus.- I.3.4.4 General comments.- I.4 Preparation of chromosome spreads.- Principle.- Protocol.- Protocol for wet slides.- Protocol for dry slides.- Remarks.- I.5 Staining techniques of chromosome spreads.- I.5.1 Classical staining using Giemsa.- Principle.- Protocol.- Remarks.- I.5.2 Orcein staining.- Protocol 1.- Protocol 2.- Remarks.- I.5.3 Acridine orange staining.- Protocol.- Remarks.- I.5.4 Propidium iodide or DAPI staining.- II Chromosome Banding Techniques.- II. 1 Introduction.- II.1.1 Chromosome organization.- II.1.2 Euchromatin.- Distribution of SINE and LINE sequences along chromosomes.- Differential arrangement of AT alignment in Q/G and R bands.- II.1.2.1 “Structural” bands.- Q bands.- G bands.- R bands.- II.1.2.2 “Dynamic” bands.- II.1.3 Code of chromosome banding techniques.- II.2 Techniques based on DNA structure.- II.2.1 QFQ bands using quinacrine mustard.- Principle.- Protocol.- Remarks.- II.2.2 GTG banding by trypsin.- Principle.- Protocol.- Remarks.- II.2.3 GAG banding by “denaturation”.- Principle.- Protocol Acid-Saline-Giemsa.- Protocol Alkaline-Saline-Giemsa.- Remarks.- II.2.4 RHG banding by thermal denaturation.- Principle.- Protocol.- Remarks.- II.2.5 T bands (terminal).- Protocol.- Remarks.- II.2.6 Bands rich in 5-methylcytocine.- II.2.6.1 Introduction.- II.2.6.2 Immunofluorescent revelation of 5-mC rich bands.- Principle.- Protocol.- Protocol of denaturation using hydrochloric acid.- Protocol of denaturation using ultraviolet lamp radiation.- Remarks.- II.3 Banding techniques based on DNA replication.- II.3.1 R or G bands by incorporation of BrdU.- Principle.- II.3.1.1 Immunoflorescent detection of BrdU incorporation in chromosomes.- Principle.- Protocol.- Remarks.- II.3.1.2 FPG staining technique (fluorochrome-photolysis-Giemsa).- Principle.- Protocol.- Remarks.- II.3.1.3 Propidium iodide staining technique.- Principle.- Protocol.- Remarks.- II.3.1.4 DAPI staining technique.- Principle.- Protocol.- II.3.1.5 Preparation of chromosomes labeled with BrdU during the second half of the S phase to produce R bands.- Protocol.- Remarks.- II.3.1.6 Preparation of chromosomes labeled with BrdU during the first half of the S phase to produce G bands.- Protocol.- Remarks.- II.3.2 Sister chromatid exchanges (SCE).- Principle.- Protocol.- Remarks.- II.3.3 Asymmetrical incorporation of BrdU.- II.4 Techniques of chromosome differentiation based on DNA base composition.- II.4.1 Treatment by 5-azacytidine or 5-azadeoxycytidine.- Principle.- Protocol.- Remarks.- II.5 Heterochromatin staining.- II.5.1 CBG bands.- Principle.- Protocol.- Remarks.- II.5.2 CT bands.- Principle.- Protocol.- II.5.3 G11 bands.- Principle.- Protocol.- Remarks.- II.5.4 DA-DAPI staining.- Principle.- Protocol.- Remarks.- II.6 Staining of nucleolar organiser regions NOR.- Protocol.- Remarks.- II.7 Techniques of sequential banding.- Principle.- Protocol of sequential Q, R, and C banding.- Protocol of sequential Q and NOR banding, or R and NOR banding.- III In Situ Hybridisation Techniques.- III.1 Introduction.- III.1.1 In situ hybridisation using radioactive probes.- III.1.2 In situ hybridisation using non radioactive probes.- III.1.3 Identification of hybridised chromosomes.- III.2 Methods.- III.2.1 Preparation of chromosome spreads.- III.2.2 DNA probe labeling.- III.2.2.1 Non radioactive labeling of long DNA probes (>1 kb).- Principle.- Protocol for nick translation labeling.- III.2.2.2 Non radioactive labelling of short DNA probes (00.25–1.5 kb).- Principle.- Protocol.- Remarks.- III.2.2.3 Radioactive labeling.- Protocol for the use of the nick translation method.- III.2.3 Pretreatment of the chromosome preparations using ribonuclease A.- Principle.- Protocol.- III.2.4 Chromosomal DNA denaturation.- Principle.- Protocol.- III.2.5 Probe preparation, labeling and denaturation.- III.2.6 In situ hybridization.- III.2.7 Posthybridisation washes.- III.2.7.1 Radioactive probes.- III.2.7.2 Non radioactive probes.- III.2.8 Hybridisation signal detection and banding.- III.2.8.1 Radioactive probes: autoradiographic detection.- Principle.- Protocol.- III.2.8.2 Non radioactives probes: immunoreaction detection.- Protocol.- III.2.9 Microscopy and photography.- III.2.9.1 Radioactive probes.- III.2.9.2 Non radioactives probes.- III.3 Remarks on other applications of in situ hybridization.- IV Methods of Germ Cells Study.- IV.1 Meiosis in male.- IV.1.1 Classical method.- Protocol.- Remarks.- IV.1.2 Synaptonemal complex method.- Principle.- Protocol.- Remarks.- IV.2 Meiosis in the mammalian female.- Principle.- Protocol.- IV.3 Study of the spermatozoa by interspecific in vitro fertilisation (insemination).- Principle.- Protocol.- Protocol of capacitation in the cold state.- Protocol of capacitation in the warm state.- Remarks.- V The Lampbrush Chromosomes of Amphibians.- V.1 Introduction: The basis of lampbrush chromosomes mapping.- V.1.1 Organisation of the lampbrush chromosomes.- V.1.2 Morphologic mapping of the lampbrush chromosomes.- V.1.2.1 Morphological variations of genetic origin.- V.1.2.2 Morphological variations of physiological origin.- V.1.3 Immunomorphological mapping.- V.1.4 Maps of the lampbrush chromosomes of Pleurodeles.- V.1.4.1 The conditions of map development.- V. 1.4.2 The intraspecific maps.- V.2 Technique for the preparation of lampbrush chromosomes for light microscopy.- V.2.1 Preparation of oocytes.- Protocol.- V.2.2 Chromosome preparation for morphological studies.- Protocol.- V.2.3 Chromosome immunolabelling.- Protocol.- V.3 Preparation of lampbrush chromosomes for electron microscopy.- V.3.1 Ultrastructural studies.- Protocol.- V.3.2 High resolution immunolabelling.- Principle.- V.3.2.1 Pre-embedding immunolabelling.- Protocol.- V.3.2.2 Post-embedding immunolabelling.- Protocol.- V.4 Preparation of mitotic chromosomes.- Protocol.- V.5 Analysis of lampbrush chromosomes.- V.5.1 Morphological markers.- V.5.1.1 Axial structures.- V.5.1.2 Loops.- V.5.2 Immunolabelling.- V.5.3 Mapping parameters.- V.5.3.1 Chromosome classification.- V.5.2.2 Chromosome orientation.- V.5.3.3 Marker localisation.- V.5.3.4 Correspondences between the lampbrush chromosome maps.- V.6 The importance of the lampbrush chromosomes.- V.6.1 Detection and analysis of chromosomal rearrangements.- V.6.2 Study of populations and evolution of chromosomes.- V.6.3 Study of the transcriptional physiology in situ.- VI Techniques for the study of Drosophila Chromosomes.- VI.1 Mitotic chromosomes.- VI.1.1 Introduction.- VI.1.2 Chromosome spreads preparation.- Principle.- Protocol.- VI.1.3 Staining and banding of mitotic chromosomes: remarks.- VI.1.4 In situ hybridization.- VI.2 Polythene chromosomes.- VI.2.1 Introduction.- VI.2.2 Structure.- VI.2.3 Reference maps.- VI.2.4 Chromosome preparation: Classical and molecular cytogenetic technique.- VI.2.4.1 Breeding conditions of Drosophila.- VI.2.4.2 Chromosome preparation: classical analysis.- Protocol.- VI.2.4.3 Chromosome preparation: in situ hybridisation.- Principle.- Protocol.- VI.2.5 In situ hybridisation.- Protocol.- VI.2.6 Chromosome observation.- VII Techniques for the stuey of Interphase Nucleus.- VII.1 Sex chromatin examination.- Principle.- Protocol.- Remarks.- VII.2 Released chromatin (Chromatin halo).- Protocol.- VII.3 In situ hybridisation of interphasic nuclei.- Principle.- Protocol.- Protocol for paraffin sections.- Protocol for nucleus and spread cells or slides with cell layers.- Remarks.- VIII Application of Flow Cytometry and Slit-Scan Fluorometry in Analysis and Sorting of Mammalian Chromosomes.- VIII.1 Introduction.- VIII.2 Principles of the flow cytometry.- VIII.2.1 Standard flow cytometry.- VIII.2.2 Slit scan fluorometry.- VIII.2.3 Flow sorting.- VIII.2.4 Computing.- VIII.3 Methods of chromosome preparation and staining.- VIII.3.1 Modified hexanediole method.- References.- Protocol.- Remarks.- VIII.3.2 TAcCaM -method.- References.- Protocol.- Remarks.- VIII.3.3 Methanol - acetic acid method.- References.- Protocol.- Remarks.- VIII.3.4 Tris/MgCl2/Triton X10000 method.- References.- Protocol.- VIII.3.5 Polyamine method.- References.- Protocol.- VIII.3.6 Modified polyamine method.- References.- Protocol.- VIII.3.7 HEPES/MgS04 method.- References.- Protocol.- VIII.3.8 Modified HEPES method.- References.- Protocol.- Remarks.- VIII.3.9 Fluorescein labelling (FITC) by in situ hybridization suspension.- References.- Protocol.- Remarks.- VIII.3.10 Dyes and lasers.- VIII.4 Measurements and evaluation in flow cytometry.- VIII.4.1 Flow karyotypes.- VIII.4.2 Data evaluation of flow karyotypes.- VIII.4.3 Slit scan measurements.- VIII.4.4 Perspectives.- VIII.4.5 Identification of the sorted chromosomes by GTG banding.- Principle.- Protocol.- Remarks.- VIII.5 In situ hybridisation to chromosomes in suspension.- Principle.- Protocol.- Remarks.- S Solutions for chromosome staining and banding techniques.- M Miscellaneous.- L Solutions for lampbrush chromosomes.- F Solutions for flow and slit scan cytometry.- H Solutions for in situ hybridization.- CM Culture media.- SS Stock solutions.- B Buffer solutions.- V Solutions for in vivo treatments.- References.