215-Lab7-Bacteria_Minipreps

215-Lab7-Bacteria_Minipreps - Lab 7 Plasmid minipreps from...

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Lab 7 – Plasmid minipreps from bacteria 7-1 Lab 7 Plasmid DNA Minipreps from Bacteria You will purify plasmid DNA from the bacterial cultures you started in the previous lab. This is the same protocol described in the lectures, although most of the explanatory notes have been deleted. While this should make the protocol easier to follow in the lab, it is very important that you understand what each step accomplishes. Remember, you are responsible for understanding each protocol used in this course. It may be helpful to refer to the flow chart for this protocol, which has been included. 1. Obtain your overnight cultures that were set up in the previous lab. 2. Tap or gently vortex each culture tube to resuspend the cells that may have settled. Pour approx. 1.5 ml of each culture to an appropriately labeled microfuge tube. Cap your microfuge tubes and spin them in the microcentrifuge at full speed (14,000 rpm) for 1 minute. 3. After spinning, pour out the clear supernatant (sup) and remove any additional liquid by using a P-200. Remove as much of the supernatant as possible without disturbing the pellet. Discard the clear supernatants into a waste container so they can be poured down the sink later. 4. Resuspend the bacterial pellet in 250 μl of Buffer P1 using the pipet tip followed by
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This note was uploaded on 10/23/2011 for the course BIO 201 taught by Professor Frenkel/cervantes during the Spring '10 term at Rutgers.

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215-Lab7-Bacteria_Minipreps - Lab 7 Plasmid minipreps from...

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