Cross-reactive antibody removal assay in antisera
This protocol describes a method for removing antibodies reactive with bacterial-encoded proteins from polyclonal antisera by co-warming the antiserum with bact
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Selection of plasmid vectors to construct expression libraries
There are three methods for detecting bound antibodies: radiochemical screening, colorimetric screening, and chemiluminescent screening, the advantages and disa
Screening experiments of large numbers of bacterial clones by hybridization methods
This method was reported by Hanahan and Meselson (1980, 1983) and is primarily used to treat bacteria transformed by plasmid cDNA libraries. A mixture of transf
BAL31 Nuclease digestion assay for generating bidirectional deletion mutants experimental
This protocol uses the nuclease BAL31 (purified from the marine bacterium Alteromonas espejiana BAL31) to produce unidirectional or bidirectional deletions in c
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
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
Experimental preparation of λ phage particles from large-scale lysates
λ phage prepared from mass cultures can be recovered from the lysate by polyethylene glycolysis (PEG) precipitation in the presence of high salt. The residual
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
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