Category: Molecular Biology

TA Cloning

As the name suggests, TA cloning involves complementation of thymine (T) and adenine (A) (complementary base pairs) that result in…

Molecular Cloning (DNA Cloning)

Molecular cloning or DNA cloning refers to a set of methods that involve in generating identical copies of a piece…

Pfu DNA Polymerase

High-fidelity thermostable DNA polymerase for error-free polymerase chain reaction. Possess 3′ to 5′ exonuclease activity for proofreading activity Produces blunt-end…

KOD DNA Polymerase

Also known as KOD HiFi DNA Polymerase High-fidelity thermostable DNA polymerase for error-free polymerase chain reaction. Originally isolated from Thermococcus…

Phusion DNA Polymerase

High-fidelity thermostable DNA polymerase for error-free polymerase chain reaction. Possess 3′ to 5′ exonuclease activity for proofreading activity Produces blunt-end…

Principle of Alkaline Lysis Method

The alkaline lysis method selectively purifies plasmid DNA from other cellular components of the bacterial cells including chromosomal DNA. Controlled…

Plasmid Isolation by Alkaline Lysis Method

Alkaline lysis method of plasmid isolation was originally developed by Birnboim & Doly (1979). In this procedure, bacteria harbouring the desired…

Protocol – Growing Large Volume of Liquid Culture of Escherichia coli for Large-Scale Plasmid Isolation

Large-scale isolation of plasmid requires a large volume of E. coli culture. Large scale plasmid isolation procedures are termed, midiprep (25 – 50 ml starting culture volume) and maxiprep (100 – 500 ml starting culture volume). A starter culture is initially prepared by inoculating a colony in a small volume (2 – 10 ml) of culture medium. Large culture volume is prepared by diluting starter culture in a ratio of 1: 100 to 1: 1000 in the growth medium.

Migration of Bromophenol Blue and Xylene Cyanol in Agarose Gel Running in TBE or TAE Electrophoresis Buffer

Bromophenol blue and Xylene cyanol are the two most commonly used tracking dyes for the analysis of DNA on agarose gel electrophoresis. These negatively charged dyes not only help in monitoring the progress of agarose gel electrophoresis but also allow easy monitoring of sample loading process onto the wells of agarose gel. Their position in relation to DNA fragments is an important information that helps to determine how far a gel must be run without accidentally letting the DNA fragments of interest run out of the gel and at the same time ensuring good resolution among different size DNA fragments.

Protocol – RNA Isolation from Tissue Using Trizol

Overview: ♦ Tissues are composed of multiple cell layers that are tightly glued together by extracellular matrix. ♦ Solubilizing all…

Protocol – RNA Isolation from Suspension Cell Culture Using Trizol

Overview: Cell lines are a very useful tool for life science research. Isolation of total RNA from cell lines is…

Protocol – RNA Isolation from Escherichia coli Using Trizol

Overview: Escherichia coli (E. coli) is one of the well characterized bacteria and is extensively used in both applied and…

Protocol – RNA Isolation from Adherent Monolayer Cell Culture Using Trizol

Overview: Cell lines are very useful tools for life science research. Isolation of total RNA from cell lines is often…

List of Monophasic Solution for RNA Extraction

TRIzol ™  Reagent (Invitrogen)  Supplied by ThermoFisher Scientific  TRI Reagent®  Supplied by MRC, Sigma Aldrich, Zymoresearch RNAzol® (Tel-Test) RNA Stat-60…