The Cryostat-Microtome

Need Microtomes for your studies? Click here A Glimpse into the History of a Cryostat A Cryostat is a versatile and high-quality machine that generates low temperatures for tissue sectioning. The word “Cryostat” originates from two separate Greek words “Kryos”, meaning cold, and “stat”, meaning stable. Sir James Dewar, a Scottish Physicist, and Chemist are […]

A Brief on The NMR Spectroscopy of Proteins

Spectroscopy is “the study of investigation and measurement of the absorption and emission of light or electromagnetic radiation by matter as a function of its wavelength and frequency”[8]. In simple terms, it’s the study of the interaction of light with a matter or substance. Depending on the nature of the interaction between the energy and […]

The Nuclear Overhauser Effect

Introduction The nuclear overhauser effect is defined as a change in the integrated intensity of one spin when the equilibrium population of another spin is perturbed by saturation or inversion. So, it is the transfer of nuclear spin polarization from one spin-active nucleus to another. However, the nuclei being irradiated should be closer to the […]

Large Scale Extraction of Bacteriophage λ DNA Using Proteinase K

Introduction and Principles Large-scale extraction of phage DNA involves digestion of viral coat proteins using a strong protease such as proteinase K followed by extraction with chloroform. This method is suitable for small-scale preparations as well.  In the experiment, dialysis of bacteriophage suspension is followed by the extraction of phage particles with chloroform: phenol, and […]

Electroporation-based CRISPR Cas9 delivery

CRISPR Cas9 is a potent and easy-to-use tool for gene editing, but the delivery of the CRISPR Cas9 system has been a challenge since it was developed. Several methods have been developed to effectively transport components of the CRISPR Cas9 system into the cells. The protocol below describes the electroporation-based delivery of Cas9 from Streptococcus […]

Polyacrylamide Gel Electrophoresis

As an Amazon Associate, ConductScience Inc. earns revenue from qualifying purchases Polyacrylamide gel electrophoresis is one of the first forms of gel electrophoresis. Polyacrylamide is a synthetic polymer that is considered the most all-around separation matrix to date due to its transparency, electrical neutrality, and adjustable pore size. The technique can be applied to the […]

Purification of Bacteriophage λ Arms using Sucrose Density Gradients

Introduction Replacement vectors or substitution vectors are prepared by removing the central filler segment of phage DNA to accommodate the fragments of foreign DNA. This procedure is known as the “preparation of λ arms.” Restriction endonucleases are used to separate the two arms of phage DNA from the central stuffer fragment. The following methods are […]

Preparation of Bacteriophage λ DNA Cleaved with two Restriction Enzymes to be used as a Cloning Vector

Introduction At each end of the central stuffer fragment, a series of restriction sites arranged in opposite directions are present in replacement vectors. For instance, in EMBL3A, the order of restriction site in the left polyclonal site is SalI-BamHI-EcoRI, and that of the right polyclonal site is EcoRI-BamHI-SalI. Digestion of this type of vector with […]

Alkaline Phosphatase Treatment of Bacteriophage λ Vector DNA

Introduction Alkaline phosphatase treatment is used to suppress the background of non-recombinants while performing restriction digestion with insertion vectors containing a single cloning site or polyclonal site using a single restriction enzyme. In this method, ligated DNA is first subjected to phenol: chloroform extraction and standard ethanol precipitation, respectively. It is then treated with restriction […]