Note: Cover -- Half Title -- Title Page -- Copyright Page -- Dedication -- Contents -- Foreword -- Preface -- Authors -- 1. DNA Manipulative and Ingratiative Techniques -- Isolation of Total Genomic DNA -- Isolation of Plant DNA Using the CTAB Method -- Introduction -- Experimental Outline -- Materials Required -- Pre-Lab Preparation -- Isolation of Plant DNA Using the SDS Method -- Introduction -- Materials Required -- Method -- Precautions -- To Isolate Plasmid DNA from Bacteria -- Introduction -- Materials Required -- Method -- Precautions -- Isolation of Chloroplasts -- Introduction -- Experimental Outline -- Materials Required -- Pre-Lab Preparation -- Method -- Results -- Precautions -- Isolation of Chloroplast DNA -- Introduction -- Experimental Outline -- Materials Required -- Pre-Lab Preparation -- Method -- Results -- Precautions -- Isolation of Yeast Genomic DNA -- Introduction -- Materials Required -- Method -- Isolation of Bacterial Genomic DNA -- Introduction -- Materials Required -- Method -- Large-Scale Isolation of Cosmid DNA -- Isolation -- Purification -- Miniprep Double-Stranded DNA Isolation -- Principle -- Isolation -- Standard Alkaline Lysis Purification -- Diatomaceous Earth-Base Purification -- Plasmid Miniprep by Birnboim Method -- Materials Required -- Plasmid Miniprep by Boiling Method -- Materials Required -- Method -- Isolation of Lambda (.) DNA -- Materials Required -- Method -- Quick Method for isolation of Lambda (.) DNA -- Materials Required -- Single-Stranded M13 DNA Isolation Using Phenol -- Method -- Biomek-Automated Modified-Eperon Isolation Procedure for Single-Stranded M13 DNA -- Method -- Isolation of Aspergillus DNA -- Materials Required -- Method -- Isolation of Aspergillus DNA by the Qiagen Method -- Materials Required -- Method -- Purification of Genomic DNA -- Introduction -- Principle -- Materials Required. , Method -- Materials Required -- Isolation of RNA from Plant Tissues -- Introduction -- Extraction of Total RNA -- Materials Required -- Method -- Precautions -- Extraction of Polysomal RNA of Nuclear Sequences -- Materials Required -- Chemicals -- Glassware and Other -- Equipment -- Isolation of Total RNA form Bacterial Cells -- Introduction -- Materials Required -- Method -- Quantitative Determination of DNA and RNA by a Spectrophotometric Method -- Introduction -- Materials Required -- Agarose Gel Electrophoresis -- Introduction -- Preparation of Agarose Gel -- Electrophoresis of DNA Fragments -- Visualization of DNA Fragments -- Materials Required -- Chemicals -- Glassware and Other -- Equipment -- Electrophoresis of DNA: Linear, Circular, and Super Coiled -- Introduction -- Experimental Outline -- Materials Required -- Method -- Precautions -- Elution of Plant DNA Fragment from Agarose Gel -- Introduction -- Materials Required -- Method -- Precautions -- Hybridization and Autoradiography of DNA -- Introduction -- Experimental Outline -- Materials Required -- Method -- Hybridization -- Autoradiography -- Precautions -- PCR in Outline -- Introduction -- Choice Between Cloning and PCR -- Experimental Outline -- Materials Required -- Method -- Precautions -- PCR Amplification of Specific Target Sequence from Genomic DNA -- PCR Consist of 3 Basic Steps -- Materials Required -- Method -- Gel Analysis -- Precautions -- Isolation of Total Plant RNA from Rice Plants and Reverse Transcriptase Polymerase Chain Reaction -- Introduction -- Nuclear Genes -- Necessary Conditions for Reaction -- Materials Required -- Pre-Lab Preparation -- Method -- Reverse Transcriptase-Polymerase Chain Reaction -- Precautions -- Isolation of Gene of Interest by PCR -- Isolation of Genomic DNA -- Materials Required -- Chemicals -- Glassware and Other -- Equipment -- Precautions. , Isolation of pUC18 plasmid from TOP10 pUC18 E. coli cells -- Materials Required -- Chemicals -- Glassware and Other -- Equipment -- Method -- Precautions -- Transformation of the Desired Bacterial Strain with Plasmid DNA -- Materials Required -- Chemicals -- Glassware and Other -- Equipment -- Pre-Lab Preparation -- Method -- Preparation of Competent Cells -- Purifying pUC18/Hind III/EcoRI Digest by Gel Elution -- Introduction -- Materials Required -- Chemicals -- Glassware and Other -- Equipment -- Precautions -- Dephosphorylation of Restriction-Enzyme-Digested Vector pUC18 -- Introduction -- Materials Required -- Method -- Precautions -- Restriction Digestion of pUC 18 and A-DNA -- Materials Required -- Chemicals -- Glassware and Other -- Equipment -- Method -- Precautions -- Enzyme Linked Immunosorbent Assay (ELISA) -- DAS-ELISA -- DAC-ELISA -- PAC-ELISA -- Materials Required -- Chemicals -- Pre-Lab Preparation -- Procedure for DAC-ELISA (Transgenic Rice) -- DAC-ELISA -- Precautions -- Electrophoretic Separation of Plasma Protein -- Materials Required -- Chemicals -- Glassware -- Equipment -- Method -- Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis -- Materials Required -- Chemicals -- Glassware and Other -- Equipment -- Pre-Lab Preparation -- Preparation of the Separating Gel -- Preparation of the Stacking Gel -- Loading and Running the Gels -- Staining and Destaining the Gels -- Method -- Production of Antibodies from Laboratory Animals -- Preparation of Immunizing Material -- Protein Solution -- Use of Adjuvants -- Alum Precipitation -- Oil in Water -- Inoculation and Bleeding of Animals -- Purification of Enzymes for Immobilization -- Equipment -- Method -- Purification of Hemoglobin -- Purification of Fungal a-Amylase -- Purification of Protease -- Enzyme Immobilization in Polyacrylamide Gel -- Equipment -- Materials Required. , Chemicals -- Materials Required -- Chemicals -- Equipment -- 2. Regulative Techniques -- Restriction Fragment Length Polymorphism (RFLP) -- Experimental Outline -- Materials and Pre-Lab Preparations -- Method -- DNA Fragment Separation on Agarose Gel -- Hybridization -- Southern Blotting Procedure -- Probe Labelling -- Autoradiography -- Amplified Fragment Length Polymorphism -- Summary of the AFLP Analysis System -- Restriction Endonucleases Digestion -- Ligation of Adapter -- Amplification Reaction -- Method -- Additional Materials Required -- Restriction Digestion of Genomic DNA -- DNA Blotting Techniques -- Precautions -- Experimental Outline -- Materials -- Pre-Lab Preparations -- Method -- Results -- Discussion -- RNA Separation of Species by Ion Exchange Column Chromatography on Methylated Albumin Kiesselguher Columns (MAK) -- Experimental Outline -- Materials -- Pre-Lab Preparations -- Method -- Preparation of Radio-Labelled Probe by Random Primer Method -- Experimental Outlines -- Materials -- Pre-Lab Preparations -- Method -- Precautions -- Detection of a Specific Fragment of Plant DNA from Restriction Digest by Hybridization with a Labelled Probe -- Experimental Outline -- Materials -- Pre-Lab Preparations -- Method -- Hybridization -- Autoradiography -- Precautions -- In Vitro Transcriptions -- Experimental Outline -- Materials -- Enzymes and Buffers -- Equipment -- Method -- To Purify RNA by Ethanol Precipitation -- Troubleshooting -- In vitro Translation of mRNA -- Experimental Outline -- Materials -- Procedure -- Prepare the mRNA by in vitro Transcription -- Prepare the Protein by in vitro Translation -- Reagents and Solutions -- Reticulocytes and Their Use in the Study of Protein Synthesis -- Experimental Outline -- Materials and Pre-lab Preparation -- Method -- Precautions -- Protein Synthesis in Intact Rabbit Reticulocytes. , Experimental Outline -- Materials and Pre-Lab Preparations -- Method -- Calculations -- Precautions -- Metabolic Labelling of Proteins and Immunoprecipitation -- Experimental Outline -- Materials and Other -- Equipment -- Method -- Western Blots -- Precautions -- Marker-Assisted Pyramiding of Leaf Rust Resistance Gene in Wheat -- Method -- PCR Reactions -- STS Marker-Based Detection of 1B/1R Translocation -- Reagents -- Method -- STS Marker-Based Detection of Lr 26 Gene -- Reagents -- Method -- Microsatellite Markers -- Advantages of Microsatellite Markers -- Applications -- Method -- Inter-Simple Sequence Repeat (ISSR) -- Reaction Mixture for ISSR -- Amplification -- Isolation of Specific Full-Length cDNAs By RT-PCR Method -- Isolation or RNAs by TRIzol Method -- Design and Synthesis of Specific Forward and Reverse Primers -- Amplification of cDNA of Interest by RT-PCR -- Purification Products by High-Speed Centrifugation of Agarose Gel Slices of PCR -- High Yield and Cleaner Elution of DNA -- Construction of Genetic Linkage Maps and the Mapping of Quantitative Trait Loci (QTLs) -- Mapmaker for the Construction of the Genetic Map, Plabstat for the Analysis of Phenotypic Data, PlaybQTL for the Interval Mapping QTL Analysis -- Transposons and Their Mutagenesis -- Site-Specific Transposon (Tn5) Mutagenesis -- Strain -- Generalized Transposon (Tn5) Mutagenesis -- Requirements -- Medium Composition -- Preparation of Antibiotic Stock Solution -- Procedure -- On 2-Dimensional Gel Electrophoresis for Analysis of Gene Expression -- Sample Preparation -- Equipment and Reagents -- Method -- Rehydration of IPG Strips -- Equipment and Reagents -- Method -- Isoelectric Focusing -- Equipment and Reagents -- Method -- Equilibration of Isoelectric Focusing Strip before Transfer to SDS-PAGE -- Equipment and Reagents -- Method -- Casting the 2-Dimensional Gel. , Equipment and Reagents.