The need to understand basic concepts in the field of experimental molecular biology is a key factor for achieving desired goals and results in this field. After post Genomic era there has been a great advancement in the field of molecular biology to exploit various signaling, biochemical, and metabolic pathways. However for the beginners in this field, it is essential that they should acquire sound knowledge of basic concepts involved in experimental molecular biology in order to master the more advanced technologies which are coming up. In this direction, we have tried to explain various basic molecular biology techniques in much palatable manner especially for undergraduate, post-graduate and fresh researchers in the form of book “An easy way to Molecular Biology Techniques”. The main aim of this book is to give an insight and full expertise to readers in their routine laboratory work in Molecular biology. Therefore, methods in this book start with some basic DNA and RNA based experimental workflow to more advanced techniques like Gene cloning, PCR, blotting etc. Suggestions by readers for improvement of this book will be highly appreciated and welcomed by the authors and these inputs used to bring subsequent revised edition of the book.

CONTENTS

Chapter 1. NUCLEIC ACID ISOLATION

1.1 Isolation of genomic DNA from Bacteria
i. Boiling method
ii. Miniprep
iii. Maxiprep

1.2 Isolation of plasmid from bacterial cell (E. coli)
i. Miniprep (Brinboim and Dolly method)
ii. Purification of DNA
iii. RNase treatment
iv. Precipitation of DNA
v. Maxiprep by alkaline lysis method

1.3 Estimation/quantification of DNA

1.4 RNA isolation from bacteria
i. RNA isolation from bacteria (E.coli)
ii. Estimation of RNA

1.5 Isolation of nucleic acids from animal tissues
i. Collection and Preservation of Biological Material
ii. DNA extraction from blood.
iii. DNA isolation from animal tissue
iv. RNA isolation from human blood using trizol method.
v. Extraction of whole cell RNA from animal tissue.

1.6 Isolation of nucleic acids from plant tissue
i. DNA extraction from plant tissue
ii. RNA isolation from plant tissue

Chapter 2. GEL ELECTROPHORESIS
2.1 Agarose Gel Electrophoresis
2.2 Sodium dodecyl sulfate - Polyacrylamide Gel Electrophoresis (SDS-PAGE)
2.3 Native PAGE
2.4 Two Dimensional Gel Electrophoresis

Chapter 3. GENERAL CLONING METHODS

3.1 Digestion of DNA by restriction enzymes.
3.2 Dephosphorylation of vector.
3.3 Ligation reactions
3.4 Purification of DNA from agarose gel.
3.5 Preparation of competent cells.
3.6 Transformation
3.7 Preparation of electro-competent cells.
3.8 Electroporation.
3.9 Blue-white screening.

Chapter 4. POLYMERASE CHAIN REACTION METHODS
4.1 PCR and its types
4.2 Reverse Transcriptase (RT-PCR)
4.3 Real time PCR (RT-PCR)
4.4 DNA sequencing.

Chapter 5. BLOTTING TECHNIQUES
5.1 Southern blotting
5.2 Western blotting