Techniques in Molecular Biology and Germplasm Conservation Techniques in Molecular Biology and Germplasm Conservation

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


Dry Heat

Red Heat



Hot Air Oven


Vaccine Bath

Serum Bath



Moist Heat Sterilisation


Non-ionising Rays

Ionising Radiations



Disinfection Techniques





Acids and Alkalis

Heavy Metals


Ethylene Oxides

Hydrogen Peroxides


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


Preservation at 70C

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



Colour Figures