Lysogenic infection assay on agar plates
Fusion proteins encoded by λ phage recombinants can be prepared by lysogenizing colonies of E. coli strain Y1089. However, preparation of lysogenic bacteria fr
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Expression of clonized genes in Escherichia coli using IPTG inducible promoter experiment
This protocol will discuss the factors affecting plasmid expression efficiency in the Troubleshooting and Optimization of Inducible Promoter Expression Proteins
Expression of cloned genes in Escherichia coli using the T7 phage promoter experiments
Tabor and Richardson in 1985 and Studier and Moffatt in 1986 proposed a new expression system utilizing the T7 phage promoter, using transcriptional signals obt
Lysogenic infection assay in liquid cultures
This method is suitable for rapid screening of immunodetectable fusion proteins against λgtll recombinant phage. However, under other conditions (e.g., optimiz
Calcium phosphate-mediated transfection of high molecular weight genomic DNA in cells
The following method is a modification of the phosphotransformation method established by Graham and Van der Eb (1973), in which high molecular weight genomic D
Calcium phosphate-mediated plasmid DNA transfection in eukaryotic cells
The calcium phosphate-mediated plasmid DNA transfection of adherent cells described in this method is an improvement on the method established by Jordan et al.
Lipofection-mediated DNA transfection assay
The following method is a modification of one studied by Mark Evans (Alexion Pharmaceuticals, New Haven, Connecticut). This experiment was derived from the next
Experiments on the purification of proteins with histidine tags by solidified Ni2+ absorption chromatography
Polyhistidine binds to a variety of transition metals and transition metal chelates, so proteins with exposed His-6 bind to cured Ni2+ resins, and target protei
Experimental extraction of λ phage DNA from mass cultures using proteinase K and SDS
DNA can be well isolated from large-scale preparations of λ phage by digestion with an effective protease (e.g., proteinase K) and subsequent phenol:chloroform