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Tag Archives: DNA isolation

Total DNA isolation protocol

The procedure is suitable for all types of tissues from wide variety of animal, blood and plant species. All DNA extraction steps are performed at weak acid pH (HEPES free acid) and optionally with hot chloroform for ‘difficult’ samples, and at room temperature. The following protocol is designed for small and large tissue samples (tissue…

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Midiprep double-stranded DNA isolation

A midi-prep double-stranded DNA isolation has been developed to generate a sufficient amount of template DNA for several Sequenase[TM] catalyzed fluorescent terminator reactions. Here, one bacterial colony which harbored the plasmid of interest is picked into 3 ml of liquid media containing ampicillin and incubated in a 37degC shaker for 8-10 hours. At this time,…

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Large scale double-stranded DNA isolation

The method used for the isolation of large scale cosmid and plasmid DNA is an unpublished modification (16) of an alkaline lysis procedure (17,18) followed by equilibrium ultracentrifugation in cesium chloride-ethidium bromide gradients (1). Briefly, cells containing the desired plasmid or cosmid are harvested by centrifugation, incubated in a lysozyme buffer, and treated with alkaline…

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