The science of agriculture is among the oldest and most intensely studied by mankind. Human intervention has led to manipulation of plant gene structure for the use of plants for the production of bio-energy, food, textiles, among other industrial uses. A sound knowledge of biotechnology and its implementation in farmer’s field through research intervention is required. This manual will provide an overview of all these aspects along with an updated review of the major plant biotechnology procedures and techniques, their impact on novel agricultural development, and crop plant improvement. This book integrates basic biotechnological methodologies along with different tools, layouts and designs of field experiments. This manual spreads overview to up-to-date agricultural practices, offering solutions to specific agricultural needs and problems from plant and crop yield. It presents and evaluates the limitations of classical methodologies and the potential of novel and emergent agriculturally related biotechnologies. The manual provides an holistic view that makes it a valuable source of information for researchers of agriculture and biotechnology as well as microbiologists, agricultural engineers, environmental biologists, environmental engineers, and environmentalists. Short exercises at the end of the chapters help to make the book suitable for course work in agricultural biotechnology, genetics, biology, biotechnology and plant science courses.
Dr. Sonia Goel, SGT University, Gurugram, Haryana
Dr. Mohinder Singh, SGT University, Gurugram, Haryana
1. Lab Safety Precautions ....................................................................... 1
2. Basic Lab Instruments ......................................................................... 5
- 2.1 Micropipettes .......................................................................................... 5
- 2.2 Analytical centrifuge .............................................................................. 6
- 2.3 Incubators ................................................................................................ 6
- 2.4 Vortex mixer ............................................................................................ 7
- 2.5 Pestle and mortar.................................................................................... 7
- 2.6 Autoclaves ............................................................................................... 8
- 2.7 Water bath ............................................................................................... 8
- 2.8 Magnetic stirrer with Hot plates ........................................................... 9
- 2.9 Forceps ..................................................................................................... 9
- 2.10 Electronic balance/digital scale ......................................................... 10
- 2.11 Bunsen Burner ...................................................................................... 10
- 2.12 Microscope ............................................................................................ 11
- 2.13 Thermometer ......................................................................................... 12
- 2.14 Laminar airflow chamber ................................................................... 12
- 2.15 Gel Doc................................................................................................... 13
- 2.16 Weighing balance machine ................................................................. 13
- 2.17 Hot air oven ........................................................................................... 14
- 2.18 pHMeter................................................................................................ 14
- 2.19 DeepRefrigerator.................................................................................. 15
- Glassware ............................................................................................ 15
- a) Beakers ................................................................................................... 15
- b) Funnels .................................................................................................. 16
- c) Volumetric flasks .................................................................................. 16
- d) Graduates Cylinders ............................................................................ 17
- e) Slides ...................................................................................................... 18
- f) Stirring rods ........................................................................................... 18
- g) Petri dish................................................................................................ 19
- h) Test tubes ............................................................................................... 19
- i) Conical flasks ........................................................................................ 19
- j) Tubes stands ......................................................................................... 20
- k) Dropper pipettes or Pasteur pipettes .................................................. 20
- l) Reagent bottles ...................................................................................... 21
- m) Trays ...................................................................................................... 21
- n) Jars ......................................................................................................... 21
- o) Micro oven ............................................................................................. 22
- p) Wash bottles .......................................................................................... 22
- q) Spatula ................................................................................................... 23
- r) Micro centrifuge racks .......................................................................... 23
- s) PCR Tubes ............................................................................................. 24
- t) Cryo tubes .............................................................................................. 24
- u) Centrifuge tubes ................................................................................... 25
3. Methods of Regents Preparations ................................................... 27
4. Extraction of Deoxyribose Nucleic Acid (DNA) from Plant Leaf Tissue ........................... 31
- Activity 1: Preparation of chemicalreagents ............................................... 32
- Activity 2: Steps of extraction of DNA .......................................................... 33
- Activity 3: Purification of DNA .................................................................... 34
5. Quantification of DNA ..................................................................... 37
- Activity 1: By gel electrophoresis utilizing EtBrfluoresence ..................... 37
- Activity 2: Spectrophotometry method ........................................................ 39
6. Polymerase Chain Reaction for DNA Amplification .................. 41
- Activity 1: Preparation of master mix ........................................................... 41
- Activity 2: Gel Based Detection of PCR products ....................................... 43
7. To Identify Importance of Morphological, Physiological and Yield Traits of Ideal Crop Plants ............................................. 47
- Activity 1: To identify quality traits and parameters of wheat crop plant ............................ 51
- Activity 2: To identify quality traits and parameters of rice crop plant ........................ 53
8. To Study Different Types of Agricultural Tools Required for Experimental Layout in the Field ................. 57
9. To Perform Field Layout for Crop Experiment ............................ 61
- Activity 1: Completely randomized design ................................................. 62
- Activity 2: Randomized complete block design ......................................... 64
- Activity 3: Split-plot design ........................................................................... 66
- Activity 4: Latin squares design.................................................................... 71
10. Chemicals and Reagents................................................................... 79
