P1 Transfer of phage clones between E. coli hosts
The yield and quality of P1 phage DNA obtained from different E. coli strains depends on the genotype of the host bacterium and the specific sequence of the exo
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Protein-protein interaction assay with GST fusion proteins
GST fusion proteins, as recombinant proteins expressed in bacteria, have been used in thousands of research projects since their introduction (Smith and johnson
Amplification and storage of mucoid libraries (amplification on filter membranes)
Applying this amplification method, the library is not prone to distortion, due to the fact that mixed colonies containing different recombinant mucilages do no
Immunoprecipitation assay to determine binding proteins
Many of the protein-protein interactions present in intact cells are maintained when cells are lysed under nondenaturing conditions. This rhopper can be used to
Experiments with two-hybrid and other two-component systems
This experiment describes experimental methods regarding two-hybrid and other two-component systems. This experiment comes from the next volume of the Laborator
Amplification and storage of mucoid libraries (amplification in liquid medium)
Do not over-amplify the mucoid library, as this will inevitably cause distortion of the original genome. Faster-growing clones will be over-presented, unstable
Ecdysone regulation induces cellular gene expression experiments
The following protocol is modified from the protocol that accompanies the Ecdysone-Induced Mammalian Expression System (Invitrogen); a similar system is sold by
Experiments on purification of radiolabeled oligonucleotides by ethanol precipitation method
If the radiolabeled oligonucleotide is to be used only as a hybridization probe, it is generally not necessary to completely remove the unadulterated radiolabel
Isolation of BAC DNA from small cultures
Small amounts of BAC DNA were prepared from 5 ml of BAC-transformed cell cultures.The DNA was prepared by alkaline lysis.The yield of BAC DNA can be as high as