Techniques in Molecular Biology and Germplasm Conservation

This book "Techniques in Molecular Biology and Germplasm Conservation" has been written to provide up-to-date and user friendly version of molecular techniques to meet the practical requirements of the students, teachers and researchers. As well as, Biotechnology, Microbiology, Biosciences, Genetics, Biochemistry, Plant Pathologists, and Plant Breeding subjects at both undergraduate and postgraduate levels. Additionally, the protocols described in this book for conservation of biological resources are of great assistance to Gene Bank Personnel and research students to maintain living organisms for short and relatively long periods.

Chapter 1: Sterilisation and Disinfection Techniques

Sterilisation Techniques

Sunlight

Dry Heat

Red Heat

Flaming

Incineration

Hot Air Oven

Pasteurisation

Vaccine Bath

Serum Bath

Inspissation

Boiling

Moist Heat Sterilisation

Radiation

Non-ionising Rays

Ionising Radiations

Filtration

Vibrations

Disinfection Techniques

Alcohols

Aldehydes

Phenols

Halogens

Acids and Alkalis

Heavy Metals

Dyes

Ethylene Oxides

Hydrogen Peroxides

Ozone

Molecular Biology and Germplasm Conservation


Chapter 2: Isolation of Microorganisms and Culture Media

Isolation of microorganisms from soil

Culture Media

Selective Media

Culture Media Fungal

Bacterial Culture Media

A. General Media

B. Semi-selective/Selective Media


Chapter 3: DNA Isolation andQuantification

DNA Isolation by Long Method

DNA Isolation by Kit Method

Bacterial DNA Isolation by Kit Method

Hybrid Method of DNA Isolation

DNA Quantification


Chapter 4: Marker Assisted Selection Techniques

PCR (Polymerase Chain Reaction)

Hybridisation Based Molecular Markers

RFLP (Restriction Fragment Length Polymorphism)

AFLP (Amplified Fragment Length Polymorphism)

PCR Based Molecular Markers

RAPD (Random Amplified Polymorphic DNA)

ISSR (Inter Simple Sequence Repeats)

CAPS (Cleaved Amplified Polymorphic Sequence)

dCAPS (Derived Cleaved Amplified Polymorphic Sequence)

SCAR (Sequence Characterized Amplified Region)

STS (Sequence Tagged Site)

Sequenced Based Markers

SSR (Simple Sequence Repeat)

SNP (Single Nucleotide Polymorphism)

EST (Expressed Sequence Tag)

DArT (Diversity Array Technology)


Chapter 5: Molecular Diagnostics Techniques

Nucleic Acid Based Techniques

Antibodies Based Techniques


Chapter 6: Conservation Techniques of Biological Resources

Short-term Methods

Frequent Sub-culturing

Storage Under Mineral Oil

Water Storage

Long-term Storage Methods

Lyophilization

Preservation at –70°C

Cryopreservation in Liquid Nitrogen

Cryopreservation in Mechanical Freezer


Chapter 7: Molecular Data Analyses

AMOVA and PCA Analyses

Population Genetic Analyses

Molecular Identification Based on Nucleotide Sequences

Phylogenetic Analysis of Nucleotide Sequences

Polymorphism Information Content (PIC) Analysis

Cluster Analysis


Annexure 1: Preparation of Chemicals/Reagents

Glossary

Index

Colour Figures