Richa Salwan, Vivek Sharma

Laboratory Methods in Microbiology and Molecular Biology (eBook, ePUB)

Methods in Molecular Microbiology

• Format: ePub

Produktbeschreibung

Laboratory Methods in Microbiology and Molecular Biology describes various microbiological, biochemical, and molecular methods employed for the characterization, identification, and analysis of actinomycetes, bacteria and fungi. The book details general guidelines, expectations, and responsibilities for good lab practices and consists of chapters that covers basic microbiological, physiological, biochemical, and molecular aspects as well as in silico analysis using various bioinformatic tools. Other topics in the book include how to make solutions, microscopy and imaging of microorganisms, sero-diagnostics, and basic concepts of phylogeny, physiology, biotechnology, soil, food, and environmental microbiology while working in laboratory. Laboratory Methods in Microbiology and Molecular Biology is an informative update to current practices and future perspectives for the field of microbial biotechnology. It aims to facilitate professors, researchers, and graduate students in monitoring the precision and accuracy of the qualitative and quantitative methods in their research.

• Involves various procedures in diverse disciplines, from microbiology to genetics, molecular biology and biochemistry
• Lists the principles and facts underlying practical applications of bacteria and fungi which have prospects in various technologies
• Includes the questions 'how' and 'why' as an explanation for novice students and researchers to modify protocols
• Facilitates students, teachers and researchers to monitor the precision and accuracy of their qualitative and quantitative methods practically

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Produktdetails

• Verlag: Elsevier Science & Techn.
• Seitenzahl: 206
• Erscheinungstermin: 13. Juni 2023
• Englisch
• ISBN-13: 9780323950794
• Artikelnr.: 68345597

Autorenporträt

Dr. Salwan is currently an Assistant Professor (Microbiology) at the College of Horticulture and Forestry (Dr. YS Parmar University of Horticulture and Forestry), Neri, Hamirpur, Himachal Pradesh, India. Dr. Salwan's research interests and contributions are on the diversity of psychrotrophic bacteria from the Western Himalayas and their utilization for industrial applications. She has also worked on the exploration of extremophiles for industrially relevant enzymes and plant beneficial microbes for agricultural benefits. She has published two books and numerous research papers in several international journals. Dr. Salwan serves as an Academic Editor for PLOS ONE and is also a recognized reviewer for several journals including MDPI Genes, MDPI Diversity, MDPI Foods, BMC Microbiology, Journal of Plant Growth Regulation, and Microbial Ecology.

Inhaltsangabe

Contents List of contributors Preface Acknowledgements Introduction
1. General guidelines, expectations, and responsibilities for inculcating
good lab practices Richa Salwan and Vivek Sharma
1. Distributing/sharing lab responsibilities 2. Awareing
students/researchers for research ethics/Authorship claims 3. Celebrations
4. Lab meetings 5. Reproducibility and data management 6. Scientific
collection/work life balance 7. Handling reagents/Solutions and visiting
fields 8. Laboratory safety 9. Personal safety 10. Preparing standard
solutions
2. Preparations of laboratory solutions Richa Salwan, Aditi Rana and Vivek
Sharma
1. Guidelines for preparing reagents, buffers, etc 2. General
considerations for working with RNA 3. General terms and description for
the preparation of reagents 4. Selection and preparation of buffer solution
References Further reading
3. Basic concepts of microbiology Richa Salwan, Aditi Rana and Vivek Sharma
1. Aim: preparation of culture media for the routine cultivation of
bacteria 2. Aim: general techniques for the isolation and cultivation of
bacteria 3. Aim: to preserve bacterial and fungal cultures by using
different methods Further reading
4. Microscopy and imaging analysis of microorganisms Richa Salwan, Aditi
Rana and Vivek Sharma
1. Aim: to prepare wet mount slide of the given sample 2. Aim: to observe
motility of bacteria using hanging drop experiment 3. Aim: preparation of
bacterial smear 4. Aim: to prepare a slide for simple staining 5. Aim: to
perform negative staining of bacteria 6. Aim: to prepare a slide for Gram's
staining 7. Aim: capsule staining by anthony staining method 8. Aim: to
perform capsule staining of the bacterial culture 9. Aim: to stain flagella
of a bacterial culture 10. Aim: to perform spore staining of the given
culture 11. Aim: perform spore staining using Dorner's method 12. Aim: to
stain the nuclear material and study the detailed morphology of the nucleus
13. Aim: to perform lactophenol cotton blue staining of the give fungal
culture 14. Aim: to perform staining of a given actinomycetes culture 15.
Aim: to perform acid-fast staining of a given culture Further reading
5. Characterization of microbes using molecular methods Richa Salwan, Aditi
Rana and Vivek Sharma
1. Morphological characterization using Biolog 2. Microbial identification
based on FAME 3. Electron microscopy 4. Molecular characterization of
bacteria Further reading
6. Soil microbiology Vivek Sharma and Richa Salwan
1. Aim: Isolation of Rhizobium from the given sample 2. Aim: To isolate
Actinomycetes from the given soil sample 3. Aim: Isolation of Azotobacter
from soil 4. Aim: To estimate soil microbial biomass by Chloroform
fumigation extraction method 5. Aim: To estimate the quantity of various
enzymes in soil samples 6. Aim: To study the effect of pH on the growth of
microbes 7. Aim: To study the effect of temperature on growth of microbes
8. Aim: To study the effect of salt concentration on growth of microbes 9.
Aim: To perform dual culture assay or direct confrontation assay References
Further reading
7. Basic experiments in food microbiology Richa Salwan, Aditi Rana and
Vivek Sharma
1. Aim: Evaluation of milk quality using methylene blue reduction test
(MBRT) assay 2. Aim: Isolation of lactic acid bacteria 3. Aim: To prepare
yogurt under laboratory conditions 4. Aim: Preparation of bread from wheat
flour 5. Aim: To prepare alcohol from grape juice under laboratory
conditions 6. Aim: To prepare sauerkraut from cabbage 7. Aim: To prepare
cheese under laboratory conditions Further reading
8. Basic experiments in microbial biochemistry Richa Salwan, Aditi Rana and
Vivek Sharma
1. Aim: Qualitative screening of bacteria for amylase production 2. Aim:
Qualitative screening of bacteria for lipase production 3. Aim: Qualitative
screening of bacteria for protease production 4. Aim: Qualitative screening
of bacteria for indole production 5. Aim: To perform methyl red (MR) test
for detection of coliforms 6. Aim: To perform Voges-Proskauer test 7. Aim:
Assessment of bacteria for utilization of sodium citrate as carbon source
8. Aim: To perform catalase test 9. Aim: Qualitative assessment of oxidase
production 10. Aim: Assessment of carbohydrate fermentation 11. Aim: To
perform oxidation-fermentation test 12. Aim: To perform nitrate reduction
test 13. Aim: Qualitative assessment of urea hydrolysis 14. Aim: To perform
triple sugar iron agar (TSIA) test Further reading
9. Analytical techniques in microbial genomics Vivek Sharma and Richa
Salwan
1. Isolation of nucleic acid (DNA and RNA) from microorganisms 2. Analysis
of nucleic acid (DNA and RNA) from microorganisms References
10. Analytical techniques in proteomics Richa Salwan and Vivek Sharma
1. Basics of protein electrophoresis 2. SDS-PAGE 3. Consumables 4.
Equipments 5. SDS-PAGE staining using Coomassie R250 or G250 6. Silver
staining 7. Chemicals 8. Equipments and per-requisite 9. Method 10.
Fluorescent silver staining 11. Chemicals 12. Equipments and per-requisite
13. Method Further reading
11. Analytical techniques in proteomics Richa Salwan and Vivek Sharma
1. Aim: Isolation and characterization of extracellular protein 2. Protein
dialysis and ultrafiltration 3. Protein fractionation and desalting using
gel filtration chromatography 4. Use of bioinformatics in planning a
protein purification Further reading
12. Expression of recombinant proteins in prokaryotic host Richa Salwan and
Vivek Sharma
1. Aim: Cloning and transformation of gene of interest in expression vector
2. Background of pET vector system and expression host 3. Equipment and
reagents for cloning and expression of genes 4. Protocol 5. Optimization of
insert: vector molar ratios Further reading
13. Sero-diagnostics in microbiology Richa Salwan and Vivek Sharma
1. Enzyme-linked immunosorbent assay (ELISA) and variants of ELISA 2.
Western blotting 3. Immunoprecipitation/agglutination assay using blood
sample 4. To perform ouchterlony double diffusion or passive double
immunodiffusion 5. To perform radial immunodiffusion assay Further reading
14. Basic tools for next-generation DNA sequencing of bacterial genome
Richa Salwan and Vivek Sharma
1. Background 2. Protocol 3. Reagents for illumina 4. Equipments 5.
Protocol 6. Bioinformatic analyses Further reading Index