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Selectivity Factor
Selectivity factor is a quantifiable measure of how efficient an antibiotic is during the process of gene selection. It measures of the capacity an antibiotic to select for transfected (resistant) cells that contain a selectable marker, while killing untransfected (sensitive) cells that do not contain a selectable marker. A selectivity factor higher than 10 is optimal. This means the concentration of antibiotic is sufficient to kill untransfected cells but not toxic enough to kill transfected cells. A selectivity factor lower than 10 means the concentration of antibiotic needed for selection is too close to the toxic concentration for the transfected cells. As a result, fewer transfected cells survive and more untransfected cells survive. In this case an alternative antibiotic should be considered. Calculating the selectivity factor The method uses a modified MTT assay. The MTT assay is a colorimetric assay used to assess cell metabolic activity. The assay is based on the reducti ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Selectable Marker
A selectable marker is a gene introduced into cell (biology), cells, especially bacteria or cells in cell culture, culture, which confers one or more traits suitable for artificial selection. They are a type of reporter gene used in laboratory microbiology, molecular biology, and genetic engineering to indicate the success of a transfection or genetic transformation, transformation or other procedure meant to introduce foreign DNA into a cell. Selectable markers are often antibiotic resistance genes: bacteria subjected to a procedure by which exogenous DNA containing an antibiotic resistance gene (usually alongside other genes of interest) has been introduced are grown on a medium containing an antibiotic, such that only those bacterial cells which have successfully taken up and gene expression, expressed the introduced genetic material, including the gene which confers antibiotic resistance, can survive and produce colony (biology), colonies. The genes encoding resistance to antibio ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
MTT Assay
The MTT assay is a colorimetric assay for assessing cell metabolic activity. NAD(P)H-dependent cellular oxidoreductase enzymes may, under defined conditions, reflect the number of viable cells present. These enzymes are capable of reducing the tetrazolium dye MTT, which is chemically 3-(4,5- di methyl thiazol-2-yl)-2,5-diphenyltetrazolium bromide, to its insoluble formazan, which has a purple color. Other closely related tetrazolium dyes including XTT, MTS and the WSTs, are used in conjunction with the intermediate electron acceptor, 1-methoxy phenazine methosulfate (PMS). With WST-1, which is cell-impermeable, reduction occurs outside the cell via plasma membrane electron transport. However, this traditionally assumed explanation is currently contended as proof has also been found of MTT reduction to formazan in lipidic cellular structures without apparent involvement of oxidoreductases. Tetrazolium dye assays can also be used to measure cytotoxicity (loss of viable cells) or ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
IC50
Half maximal inhibitory concentration (IC50) is a measure of the potency of a substance in inhibiting a specific biological or biochemical function. IC50 is a quantitative measure that indicates how much of a particular inhibitory substance (e.g. drug) is needed to inhibit, ''in vitro'', a given biological process or biological component by 50%. The biological component could be an enzyme, cell, cell receptor or microbe. IC50 values are typically expressed as molar concentration. IC50 is commonly used as a measure of antagonist drug potency in pharmacological research. IC50 is comparable to other measures of potency, such as EC50 for excitatory drugs. EC50 represents the dose or plasma concentration required for obtaining 50% of a maximum effect ''in vivo''. IC50 can be determined with functional assays or with competition binding assays. Sometimes, IC50 values are converted to the pIC50 scale. :\ce = -\log_ \ce Due to the minus sign, higher values of pIC50 indicate ex ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
Transfection
Transfection is the process of deliberately introducing naked or purified nucleic acids into eukaryotic cells. It may also refer to other methods and cell types, although other terms are often preferred: " transformation" is typically used to describe non-viral DNA transfer in bacteria and non-animal eukaryotic cells, including plant cells. In animal cells, transfection is the preferred term, as the term "transformation" is also used to refer to a cell's progression to a cancerous state (carcinogenesis). Transduction is often used to describe virus-mediated gene transfer into prokaryotic cells. The word ''transfection'' is a portmanteau of the prefix ''trans-'' and the word "infection." Genetic material (such as supercoiled plasmid DNA or siRNA constructs), may be transfected. Transfection of animal cells typically involves opening transient pores or "holes" in the cell membrane to allow the uptake of material. Transfection can be carried out using calcium phosphate (i.e. ... [...More Info...] [...Related Items...] OR: [Wikipedia] [Google] [Baidu] |
