Molecular Biology. A Practical Guide - Velikov V.A. 2013



Introduction

Isolation of Total DNA from Cells

Isolation of DNA from Bacteria

Isolation of DNA from Blood Leukocytes

Isolation of DNA from Plant Tissues

DNA Concentration by Alcohol Precipitation

Spectrophotometry of DNA, RNA, and protein preparations

Spectrophotometric determination of DNA concentration

Spectrophotometry of DNA, RNA, and Protein Preparations

Determination of Protein Concentration via Intrinsic Fluorescence

Nucleic Acid Electrophoresis

Preparation of Agarose Gel

DNA Gel Electrophoresis

Isolation of Plasmid and Phage DNA

Mini-preparation of Plasmid DNA (Miniprep)

Large-Scale Isolation of Plasmid DNA (Maxiprep)

Isolation of the M13 Phage Replicative Form (RF)

Isolation of M13 Single-Stranded Phage DNA

DNA Restriction

Restriction of Plasmid and Phage DNA

Restriction of Eukaryotic Genomic DNA

PCR

Primer Design

Performing DNA PCR amplification

Primer design and PCR parameter calculation using specialized software

Gene Cloning

Preparation of Vector DNA and Target Gene

DNA Ligation of Vector and Gene

Gene cloning

Restriction mapping of a DNA insert in a plasmid

Bacterial Transformation

Preparation of Competent E. coli Cells

Transformation of E. coli

Preparation of Electrocompetent E. coli Cells

Electroporation of E. coli

Isolation of Recombinant Protein

Induced Expression of Cloned Genes

Extraction of Protein from the Periplasm of E. coli Cells

Isolation of recombinant protein

Dialysis of the protein preparation

Determination of Protein Concentration

Bradford Assay

Lowry Method

Protein Concentration by TCA Precipitation

Protein Electrophoresis

Preparation of Solutions and Casting of PAGE

Performing Protein Electrophoresis

Coomassie R-250 Protein Staining

Calculations in Protein Work

Translation of Nucleotide Sequences

Calculations in Protein Research

Prediction of Nucleotide Sequence from Amino Acid Sequence

Analysis of Protein Amino Acid Sequences

Recommended Reading

Appendix