Establishment of randomized overlapping DNA insertion libraries
The following protocol describes the starting steps of birdshot sequencing, from shearing of target DNA to construction of a DNA fragment library with M13 phage
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Agarose gel electrophoresis of DNA
DNA electrophoresis can be used to (1) separate DNA fragments of different sizes, (2) identify target DNA fragments, and (3) purify and recover DNA fragments.
Ligation of plasmid and target DNA in low melting point agarose experiment
The most time-consuming step of the plasmid cloning operation is the purification of the desired sized fragments of exogenous and plasmid DNA by electrophoresis
Screening of bacterial colonies with X-gal and IPTG (alpha complementary)
The chromogenic substrate X-gal can be mixed with bacterial cultures, mixed with solubilized top agar and spread on selective culture plates. This experiment is
λ Experiments on spread plate culture of phages
Phages originate from the infection of a single bacterium by a single phage particle. The daughter virus particles synthesized after the first infection adsorb
Preparation of cytoplasmic S-100 components
The contents of a cell contained within the cell membrane. In eukaryotic cells it refers to the portion of the cell membrane outside the inner core that contain
Hybridization of bacterial DNA on filter membranes
This protocol describes how to hybridize radiolabeled probes to DNA of transforming bacteria immobilized on filter membranes, and methods for recovering clones
Preparation of λ phage proto-species assay by plate lysis and elution
The preparation of λ phage progenitors derived from a single phage spot is usually done by two methods: (i) plate lysis, which is a method of phage proli
DNA binding protein assay
This experiment was used to detect sequence-specific DNA-binding proteins in crude extracts.