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ZOO731 : Advanced Analytical Techniques

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Course Info

Course Category

Biotechnology

Course Level

Graduate

Credit Hours

3

Pre-requisites

N/A

Instructor

Prof. Dr. Muhammad Arshad
Ph.D. (Molecular Immunology)
Medical University of Vienna, Austria

Course Contents

 

Differential Staining Techniques:

Introduction & need of staining

Common biological stains

Stainability of tissues

General staining procedure

Types of staining techniques

Simple staining

Differential staining

Acid-fast staining

Ziehl-Neelson staining

Albert staining

Silver staining

Haematoxylin and eosin (H&E) staining

 

Animal preservation techniques:

Introduction and Objectives

Preservation techniques

Methods

Factors affecting preservation

Applications


Sampling techniques:

Population & its types

What is a sample?

Sampling criteria

Sampling size

Purpose of sampling

Types of sampling techniques:

Probability sampling:

Simple random

Systematic

Stratified

Cluster

Non-probability:

Accidental

Quota

Purposive


Microtomy:

Introduction

Types

Method

Fixing

Processing

Embedding

Sectioning

Staining

Applications

 

Cryopreservation:

Introduction & history

Cryopreservation agents

Natural Cryopreservation

Temperature

Risk factors

Methods of Cryopreservation

Applications

 

Cell fractionation

Introduction

Method

Phases of cell fractionation

Homogenization

Filtration

Purification

Applications

 

Agarose gel electrophoresis

Introduction

What is agarose?

Properties of AGE

Requirements for AGE

Procedure

Casting of gel

Loading of samples

Electrophoresis

Staining & visualization

Factors affecting AGE

Applications of AGE

 

Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE):     

Introduction

Principle

Requirements for SDS-PAGE

Procedure

Factors affecting SDS-PAGE

Applications SDS-PAGE

 

HPLC (High performance liquid chromatography:

Introduction

Principle

Types

Isocratic & gradient elution

Factors affecting HPLC

Applications of HPLC

 

Gas chromatography:

Introduction

Principle

Method

Data analysis

Advantage & disadvantage

Applications in real world

 

Size exclusion chromatography

Introduction

Basic principle

method

Factors affecting filtration

Absolute size exclusion chromatography

Advantages & disadvantages

Applications

 

Column chromatography:

Introduction

Basic principle

Components

Method

Applications

 

PCR:

Introduction

Principle

Components

Procedure

Types

Advantages & Disadvantages

Applications

 

DNA sequencing:

Introduction

Structure of DNA

Methods

Basic Methods

Maxam-Gilbert sequencing

Chain-termination sequencing

Advance methods

Shortgun sequencing

Bridge PCR

Uses of sequencing

 

Protein sequencing:

Introduction

Structure of Proteins

Method

Uses of sequencing

 

Estimation of proteins:

Introduction

Determination of Overall   Protein Concentration (methods):

Kjeldahl method

Methods using UV-visible spectroscopy

Advantages & disadvantages

Protein Purification

Characterization of proteins

Applications

 

Estimation of lipids:

Introduction

Structure of lipids

Methods

Applications

 

Estimation of DNA:

Introduction

Structure of DNA

Methods

Applications

 

Northern blotting:

Introduction

Northern blotting Vs other blotting techniques

Procedure

Advantage & disadvantage

Reverse northern blotting

Applications

 

Western Blotting:

Introduction

Western blotting Vs other blotting techniques

Components

Protocol

Applications

 

Southern blotting:

Introduction

Principle

Procedure

How to label probe

Detection methods

Applications

 

Biosensor:

Definition

Components

Principle

Basic characters

Types of biosensor

Applications of biosensors

 

 

DNA Barcoding:

Introduction,

DNA barcode- A tool of genetic analysis

Mitochondrial genome

Ideal gene for DNA barcoding (Choice of Locus)

Procedure & steps of DNA barcoding 

Applications of Barcoding

Advantages & disadvantages of DNA barcoding

 

ELISA:

Introduction

Basic principle of ELISA technique

Types:

Direct ELISA

Indirect ELISA

Sandwich ELISA

Competitive ELISA

Applications

 

Flow cytometry:

Introduction

Principle of flow cytometer

Components

Data analysis

Fluorescence activated cell sorting (FACS)

Measurable parameters

Applications

 

 

Immunofluorescence:

Introduction

Principles of fluorescence

Types of fluorescence:

Direct

Indirect

Uses of fluorescence:

Fluorescent Microscopy

Fluorescent conjugates

Limitations of immunofluorescence

Applications

 

Immunoprecipitation:

Definition

Principle

Methods:

Direct

Indirect

Types:

Immunoprecipitation (IP)

Co- Immunoprecipitation (CoIP)

Chromatin Immunoprecipitation (ChIP)

RNA Immunoprecipitation (RIP)

Procedure

Applications

 

Luciferase reporter assay:

What is luciferase assay?

Principle of luciferase assay

Method

Reporter gene

Applications

 

Radioimmunoassay (RIA):

Introduction

How RIA works

Procedure

Uses of RIA

Advantages & disadvantages

 

Immunohistochemistry:

Introduction

Protocol

Sample preparation

Sample labelling

IHC reporter

Detection of target antigen

Counterstaining

Sealing the stained sample

Visualization of samples

Applications

 

Fluorescence in situ hybridization (FISH):

Introduction 

How FISH works?

Method

Preparation and hybridization process

Applications

 

Chemiluminescence:

Introduction

Principle

Method

Draw backs

Applications