Essential Laboratory Techniques of Plant Biotechnology, Biochemistry and Physiology
Debanjana Saha, Udit Nandan Mishra, Ankita Mohanty, Rajesh Kumar Singhal
Background
The Laboratory Manual of Essential Techniques in Plant Biotechnology, Biochemistry, and Physiology is a comprehensive, hands-on guide designed for students, educators, and researchers in plant science. It bridges theoretical concepts with practical skills, offering detailed protocols, clear objectives, and step-by-step instructions for a wide range of laboratory techniques. The manual covers core areas such as DNA extraction, PCR, enzymatic assays, metabolic profiling, and physiological experiments related to photosynthesis, transpiration, and stress responses. By integrating plant biotechnology with biochemical and physiological approaches, it highlights applications in crop improvement, stress adaptation, and sustainable agriculture. This manual not only supports learning but also fosters critical thinking, data analysis, and experimental design, making it a vital resource for experiential learning and scientific growth in the plant sciences.
Contents of the book
SECTION-A
- Laboratory setup and requirements for plant tissue culture
- Sterilization procedure for plant tissue culture
- Preparation of MS nutrient media
- Direct and indirect organogenesis of plant tissue culture
- Embryo culture
- Anther culture
- Somatic embryogenesis
- Cell suspension
- Isolation of plant genomic DNA by CTAB method
- RNA isolation by TRIzol reagent
- PCR analysis
- Agarose gel electrophoresis
- Gene cloning
- Gene transformation in plant leaf discs mediated by agrobacterium
- Gene transformation in plants mediated by particle bombardment
- Southern blotting
- Northern blotting
- Western blotting
- RT-PCR analysis
- cDNA library preparation
SECTION-B
- Quantitative estimation of titratable acidity using titration
- Preparation of buffer solutions of varying pH
- Estimation of total soluble sugars (total carbohydrates)
- Estimation of reducing sugars
- Estimation of starch by anthrone reagent
- Estimation of nitrogen by Micro-Kjeldahl Method
- Estimation of protein by Biuret Method
- Estimation of protein by Lowry's Method
- Estimation of protein by Uv-Absorption (A280) Method
- Separation of plant pigment through paper chromatography
- Action of salivary amylase on storage polysaccharide under varying pH and temperature range
- Polyacrylamide-Sodium Dodecyl Sulphate Gel Electrophoresis (SDS-PAGE) of proteins
- Extraction of total lipids
- Estimation of oil or crude fat content in oilseeds
- Isolation and estimation of free fatty acids
- Determination of free fatty acids or acid value of an oil
- Determination of iodine value of an oil
- Determination of saponification value of fat/oil
- Extraction of enzymes
- Assay of amylase enzymes
- Assay of lipase enzyme
- Estimation of chlorophyll
- Estimation of oxalic acid
- Antioxidant enzyme assay