Basic Life Science Methods

A Laboratory Manual for Students and Researchers

1st Edition - September 11, 2022
Authors: Javeed Ahmad Tantray, Nighat Un Nissa, Rasy Fayaz Choh Wani, Sheikh Mansoor Shafi
Language: English
Paperback ISBN:
9 7 8 - 0 - 4 4 3 - 1 9 1 7 4 - 9
eBook ISBN:
9 7 8 - 0 - 4 4 3 - 1 9 1 7 5 - 6

Basic Life Science Methods: A Laboratory Manual for Students and Researchers presents forty of the most executed life science assays. The authors use a consistent structure to cov… Read more

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Basic Life Science Methods: A Laboratory Manual for Students and Researchers presents forty of the most executed life science assays. The authors use a consistent structure to cover the preparation, execution and analysis of data from each method. Assays include estimation of cholesterol fractions, C-Reactive Protein, Genomic DNA isolation, Agarose Gel Electrophoresis, RT-PCR, DNA solution preparation, how to design primers, and enzyme-linked immunosorbent assay (ELISA). This book provides a complete reference containing step-by-step instructions on how to run life science assays. Laboratory staff can also benefit of the book as a training resource.

Cover image
Title page
Table of Contents
Copyright
List of figures
List of tables
About the authors
Foreword
Foreword
Introduction
Chapter 1. Calculations of molarity and normality
Molarity (M)
Normality (N)
Chapter 2. Glassware washing and sterilization techniques
Principle
Washing of glasswares is done by two main methods
General procedure for cleaning of glasswares by detergent
General procedure for cleaning of glasswares by chromic acid
Preparation of 5% chromic acid
Cleaning by chromic acid
Types of sterilization
Observation and results
Chapter 3. pH meter: Its use and calibration
Structure
Calibration
Observation and result
Chapter 4. Principle and working of autoclave
Conditions for autoclave sterilization
Chapter 5. Principle and working of centrifuge
Rate of sedimentation
Sedimentation time
Parts of centrifuge
Relative centrifugal force
Types of centrifuges
Type of centrifugations
Chapter 6. Principle and working of laminar
Parts of a laminar air flow chamber (Fig. 6.1)
Advantages of a laminar air flow chamber
Disadvantages of a laminar air flow chamber
Applications of a laminar air flow chamber
When employing a laminar air flow chamber, there are a few things to keep in mind
Chapter 7. Estimation of total cholesterol (Flegg, 1972)
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Expected outcomes
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Expected outcomes
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Expected outcomes
Advantages
Limitations
Safety considerations and standards
Chapter 8. Estimation of creatinine
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Expected outcomes
Advantages
Limitations
Safety considerations and standards
Alternative methods/procedures
Chapter 9. Estimation of blood urea
Materials and equipment
Step-by-step method details
Expected outcomes
Advantages
Limitations
Safety considerations and standards
Alternative methods/procedures
Chapter 10. Estimation of blood glucose by GOD—POD method
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Expected outcomes
Normal range blood
Advantages
Limitations
Safety considerations and standards
Alternative methods/procedures
Chapter 11. Determination of albumin and globulin in plasma
Materials and equipment (materials and reagents)
Chapter 12. Quantitative determination of homocysteine
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Expected outcomes
Advantages
Limitations
Safety considerations and standards
Chapter 13. Estimation of serum bilirubin
Reagents required
Materials required
Reagent preparation
Assay procedure
Assay procedure
Chapter 14. Estimation of diagnostic enzyme ALT
Reagents
Materials required
Regent preparation
Specimen
Assay procedure
Assay procedure 2
Calculations
Quality control
Performance characteristics
Temperature conversion factors
Chapter 15. Estimation of diagnostic enzyme AST
Reagents
Materials required
Regent preparation
Specimen
Assay procedure
Sample/control
Assay procedure 2
Sample/control
Calculations
Quality control
Performance characteristics
Chapter 16. Protein estimation by Lowry's method
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Expected outcomes
Advantages
Limitations
Safety considerations and standards
Alternative methods/procedures
Chapter 17. Estimation of reducing sugar by using dinitro salicylic acid method
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Expected outcomes
Advantages
Limitations
Safety considerations and standards
Alternative methods/procedures
Chapter 18. To perform quantitative determination of total protein in serum/plasma
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Expected outcomes
Advantages
Limitations
Safety considerations and standards
Alternative methods/procedures
Chapter 19. Enzyme activity using starch assay
Requirements
Preparation
Procedure
Observation and result
Determination of enzyme activity
Chapter 20. To estimate protein by Bradford assay
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Expected outcomes
Advantages
Limitations
Safety considerations and standards
Chapter 21. To identify a protein sample using SDS-PAGE
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Expected outcomes
Advantages
Limitations
Safety considerations and standards
Chapter 22. Genomic DNA isolation from whole blood
Principle
Materials and equipment (materials and reagents) (Tables 22.1–22.3)
Step-by-step method details (experimental procedure)
Expected outcomes
Advantages
Limitations
Safety considerations and standards
Chapter 23. PCR for gene amplification
Materials and equipment
Expected outcomes
Advantages
Limitations
Safety considerations and standards
Chapter 24. Agarose gel electrophoresis
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Expected outcomes
Advantages
Limitations
Safety considerations and standards
Chapter 25. RNA isolation from human tissue
Some important precautions
Requirements
Protocol
Optimized protocol for the extraction of RNA isolation from blood
Chapter 26. Real-time polymerase chain reaction
Types of real-time quantification
Materials and equipment
Step-by-step method details
Relative gene expression studies
Expected outcomes
Advantages
Limitations
Safety considerations and standards
Chapter 27. Preparation of competent cells by CaCl2 treatment
Principle
Materials required
Equipment
Principle
Preparation of media or solution
Procedure
Methods
Heat shock transformation of Escherichia coli
Methods
Chapter 28. RNA isolation from plant tissues
DNase treatment
cDNA synthesis
Chapter 29. DNA isolation from the bacterial culture broth
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Expected outcomes
Advantages
Limitations
Safety considerations and standards
Chapter 30. DNA isolation from fungal mycelium
Introduction
Procedure or steps
Precautions
Chapter 31. Quantification of the DNA using spectrophotometer
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Expected outcomes
Advantages
Limitations
Safety considerations and standards
Chapter 32. Formaldehyde gel electrophoresis of the isolated RNA sample
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Expected outcomes
Advantages
Limitations
Safety considerations and standards
Chapter 33. Paper chromatography of different amino acid
Materials required for setup of technique
Experimental procedure
Biosafety considerations and laboratory standards
The experimental outcome
Advantages
Limitations
Chapter 34. Separation of plant pigments using column chromatography
Basic principle behind separation of biomolecules
Materials and equipment (materials and reagents)
Chemical solutions used for column chromatography
Step-by-step procedure
Expected outcomes
Advantages of column chromatography
Main limitations
Safety concerns and experimental standards
Alternative methods/procedures
Chapter 35. Separation of plant pigment by thin-layer chromatography
Materials required
Reagents
Procedure
Observation and results
Chapter 36. Chromatin immunoprecipitation protocol
Materials
Procedure
Precautions
Chapter 37. Types of media
Chapter 38. Preparation of nutrient agar media
Principle
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Expected outcomes
Advantages
Limitations
Safety considerations and standards
Chapter 39. Serial dilution of the bacterial cells
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Advantages
Limitations
Safety considerations and standards
Chapter 40. Spread plate method of the bacterial cells
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Expected outcomes
Advantages
Limitations
Safety considerations and standards
Chapter 41. Streak plate method
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Expected outcomes
Advantages
Limitations
Safety considerations and standards
Chapter 42. Identified bacterial colony in culture broth
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Expected outcomes
Advantages
Limitations
Safety considerations and standards
Chapter 43. Pour plate method for bacterial colony counting
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Expected outcomes
Advantages
Limitations
Safety considerations and standards
Chapter 44. Gram staining of bacteria
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Expected outcomes
Advantages
Limitations
Safety considerations and standards
Chapter 45. Casein hydrolysis test
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Expected outcomes
Advantages
Limitations
Safety considerations and standards
Chapter 46. Antibiotic resistance test by agar well diffusion method
Materials and equipment (materials and reagents)
Step-by-step method details (experimental procedure)
Expected outcomes
Advantages
Limitations
Index

Javeed Ahmad Tantray

Dr. Javeed Ahmad Tantray’s current research focus is on Human cardiovascular diseases obtained his Ph. D from Osmania University in Hyderabad, India. Dr. Tantray has done Post Doc studies with the Department of Zoology, University of Kashmir, receiving funding from the UGC-CSIR. He published many National and International peer-reviewed research articles. Dr. Tantray has published in high impact factor journals such as the International Journal of Cardiology, and Saudi journal of biological science. He has guided 50, M.Sc / and graduate students projects. He has received Junior scientist awards both at national and international level. He received the Best Abstract Award at the 4th Asia Pacific CardioMetabolic Syndrome Congress held by the Korean Society of CardioMetabolic Syndrome,with an award amount of USD 500. Dr. Tantray has organized a national seminar on "Cancer Bio-markers" and a one-day national workshop on “Assembly and usage of Foldscope”. Dr. Tantray is currently an Assistant Professor in the Department of Zoology, School of life sciences central university of Kashmir (J&K).

Affiliations and expertise

Assistant Professor, Department of Zoology, Central University of Kashmir, India

Nighat Un Nissa

Nighat Un Nissa has completed her M.Sc. and M.Phil. From Department of zoology, University of Kashmir. Ms. Nissa is currently pursuing her Ph.D. in Zoology and is working on Fish parasites and their impact on pollution. She has published her research work in National and International journals. She was awarded Gold Medal for poster presentation in Animal science congress held by Department of Zoology, University of Kashmir.

Affiliations and expertise

Assistant Professor, Department of Zoology, University of Kashmir, India

Rasy Fayaz Choh Wani

Rasy Fayaz Choh Wani PhD has completed her M.Sc. degree from Kashmir University. Ms. Rasy is currently pursuing Ph.D. She has published her research work in many reputed journals. She has presented her different papers in different international and national conferences. She is currently on research topic “Isolation and characterization of pathogenic bacterial microflora associated with cultured Cyprinus carpio in Kashmir Valley”.

Affiliations and expertise

Research Scholar, Department of Zoology, University of Kashmir, India

Sheikh Mansoor Shafi

Dr. Sheikh Mansoor Shafi currently serves as a postdoctoral researcher at the Phenomics Laboratory within Jeju National University in South Korea. His primary research focuses on high-throughput phenomics, remote sensing and precision agriculture. Prior to this role, he held positions as a Postdoc Research Associate at SKIMS Srinagar and as a Project Associate-I at CSIR-Indian Institute of Integrative Medicine Jammu (IIIM). Throughout his academic journey, Dr. Mansoor has had the opportunity to work in reputed laboratories, including the Transcriptomics Labs at SKUAST Kashmir and the Department of Biotechnology and Pathology at Central Institute of Temperate Horticulture (CITH) Rangreth Srinagar. His educational background comprises a BSc degree from SP College of Science (University of Kashmir), an MSc degree from HNBGU Uttarakhand, and a Ph.D. degree from Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu. Dr. Mansoor has been dedicated to contributing significantly to his field and has displayed a strong commitment to becoming a highly accomplished scientist. His expertise spans various areas, including biochemistry, molecular biology, and genetics. Dr. Mansoor's accomplishments are notable, including over more than 50 research and review papers published in esteemed journals such as Frontiers, Plos One, Scientific Reports, Journal of Fungi (JoF), Planta, Chemosphere, JEMA and Molecules etc. He has also authored 5 books and more than ten book chapters, and holds credit for filing two patent applications. He has showcased his knowledge through guest lectures on topics like sequence assembly, alignment, submission tools, and phylogenetic analysis. He has received practical training in advanced techniques in modern biology and related fields. Dr. Mansoor has been recognized with numerous national and international accolades, including prestigious awards such as Best Thesis, Young Scientist, and recognition for his exceptional oral and poster presentations. He holds membership in a significant big grant (BIRAC) awarded by the Department of Biotechnology (DBT) in India. In addition to his accomplishments, Dr. Mansoor plays an active role as a Review Editor for Frontiers in Ethnopharmacology, and he also contributes as a reviewer for esteemed journals like 3 Biotech, Cell Reports, Frontiers, Plos One, and JARMAP. His unwavering dedication, profound expertise, and active involvement underscore his unwavering commitment to advancing scientific knowledge in his field.

Affiliations and expertise

Postdoctoral Researcher, Phenomics Laboratory, Department of Plant Resources and Environment, Jeju National University, South Korea