BIO 480W-Genomic DNA Isolation Protocol (Lab 7)(2)

BIO 480W-Genomic DNA Isolation Protocol (Lab 7)(2) - BIO...

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BIO 480W Dr. Gaillard Spring 2009 Lab 7 Isolation of Chromosomal DNA from Bacillus licheniformis (using the QIAGEN Genomic DNA Isolation Protocol) Procedure : 1. Pellet 8 ml of the bacteria by centrifuging at 5000 RPM for 6 minutes. 2. Taking care not to disturb the pellet, remove the supernatant and discard it. 3. Add 3.5 ml of Buffer B1 (containing RNase A) to the pellet and resuspend the pellet by vortexing at top speed. Continue vortexing the bacteria until a homogenous suspension is obtained (no visible clumps of bacteria). 4. Add 80 μ l of lysozyme stock solution and 100 μ l of Proteinase K stock solution to the bacterial suspension. Incubate the suspension at 37°C for 45 minutes. During incubation, lysozyme digests the bacterial cell wall, while the detergents in Buffer B1 break open the bacterial cell membrane. RNase A and Proteinase K digest intracellular RNA and protein, respectively. 5.
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This note was uploaded on 04/02/2009 for the course BIO 480W taught by Professor Gaillard during the Spring '09 term at Sam Houston State University.

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BIO 480W-Genomic DNA Isolation Protocol (Lab 7)(2) - BIO...

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