BIT410Lab2

BIT410Lab2 - E-coli Bacterial Pellet pEGFP-N1 Plasmid...

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E-coli Bacterial Pellet pEGFP-N1 Plasmid Purification 2/4/10 BIT 410, 202 Purpose: Isolate plasmid DNA from e-coli for the purpose of cloning. In this lab pEGFP-N1 was purified however later labs will also require a second plasmid, pET41a to be purified. Alkaline Lysis followed by anion exchange chromatography was used for purification from chromosomal DNA, RNA and cell proteins. Quantification of the final plasmid solution was obtained using Nanodrop. Materials and solutions were supplied from the QIAfilter Plasmid Maxi Kit. Materials and Methods: Resuspend E-coli bacterial pellet in P1 Buffer by vortexing or repeated pipetting. No clumps should be in solution. Add Buffer P2 and mix gently by inverting tube then let incubate at room temperature for 5 minutes. During the incubation period prepare the QIAfilter Cartridge by screwing the cap on the end nozzle. Once incubation time is over add Buffer P3 to lysate and mix gently. Pour lysate into barrel of the QIAfilter Cartridge and let incubate at room
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This note was uploaded on 08/22/2010 for the course BIT 410 taught by Professor Lady during the Spring '10 term at N.C. State.

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BIT410Lab2 - E-coli Bacterial Pellet pEGFP-N1 Plasmid...

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