9 Laboratory Equipment - Electrophoresis www.alphalabs.co.uk ■ Additional Products and Information ■ Technical Specifications ■ Supporting Literature and videos ■ Improved navigation and functionality n email: export@alphalabs.co.uk Technical Specifications Unit dimensions (W x L x H) 31 x 34 x 9cm Max. sample capacity 20 Buffer volume 1000ml Recommended Running Conditions 25V/300mA for 1 hour Comet Assay Tank Catalogue Number Description EL1920 Comet assay tank for SCGE for up to 20 slides Each tank’s robust construction from ebony acrylic ensures that cells remain free of exposure to background light and DNA damage during electrophoresis, while a cooled central platform provides a convenient surface for slide preparation and control of slide temperature during the assay. Following electrophoresis DNA damage may be measured using Comet Assay scoring software. n For Single Cell Gel Electrophoresis n Minimise exposure to light and reduce background DNA damage n High efficiency cooling for enhanced resolution Single Cell Gel Electrophoresis Initially back in the 1980’s SCGE was used to quantify double-stranded DNA breakages in single cells exposed to irradiation. The technique has since been adapted to analyse specific DNA lesions and repair processes. Cells are exposed to a genotoxic insult, such as ionising radiation, which results in strand breakage of supercoiled duplex DNA, thereby reducing the size of the large genomic DNA molecules. Following this treatment, cells are suspended within low melting point agarose which is smeared on to a microscope slide to create a thin layer in which the cells are embedded. Cellular protein is then removed by lysis in detergent, and the alkaline conditions promote the unwinding of the DNA in readiness for electrophoresis. The DNA is electrophoresed, stained and then analysed using software. The electrophoresis draws out the DNA fragments and the genomic DNA takes on the appearance of a comet as its negatively charged broken ends and fragments migrate towards the anode during electrophoresis. The head of the comet is mainly composed of intact genomic DNA, whereas any fragmented or damaged DNA is concentrated within and towards the tail. Microscopy imaging is used to measure DNA fluorescence upon staining.
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