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March 29, 2021

Laboratory Techniques

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 ultimately removal of CsCl. The
Laboratory Techniques

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 pyogenes coupled with synthetic and
SDS-Polyacrylamide Gel Electrophoresis at Neutral pH (NuPAGE)
Laboratory Techniques

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 analysis of proteins and nucleic
Laboratory Techniques

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 "preparation of λ arms." Restriction endonucleases are used to separate the two arms of phage DNA from the central stuffer fragment. Following methods are used to purify bacteriophage λ arms: Sucrose