DNAase I hypersensitive site mapping
DNAase I hypersensitive site mapping is frequently used to localize regulatory regions of eukaryotic genes. For reasons that are not yet understood, genes carry
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DEAE-dextran-mediated high-efficiency transfection assay
Two forms of the classical DEAE-dextran transfection method are introduced here. The first one (the main option), in which the cells are exposed to high concent
Screening of unamplified mucoid libraries by hybridization (filter film photocopying)
Thick bacterial colonies transformed by mucilage can be screened by hybridization, a method derived from the large-scale screening of plasmid-transformed coloni
Experiments with tetracycline as a regulator of inducible gene expression
The following protocol is divided into 3 stages: stable transfection of fibroblasts with pTet-tTAk, stable transfection of NIH-3T3 cells inducible to express tT
Conjugate transcription profiling experiments
Nuclear contig analysis is used to determine whether a gene is transcriptionally regulated in a given clone. Theoretically, nuclear contig analysis should be pe
Determination of chloramphenicol acyltransferase content by thin-layer chromatography experiment
In this protocol, we list three different assays that can be used to determine CATase activity expressed after transfection of mammalian cells with the reporter
Determination of luciferase in mammalian cell extracts
The assay of luciferase in mammalian cell extracts can be used to (1) measure the amount of luciferase in animal tissues, and (2) measure other enzymatic activi
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