Plant DNA Extraction I

We were given 5 plant samples (I got the one from Presidio) and inserted each sample into a 1.5mL microcentrifuge tube along with 3 sterile steel balls. We then placed those in a rack and attached it to this wooden block and broke down the cell walls using a reciprocating saw for 40s – the final product looked like a green powder. We centrifuged this so that the green powder would collect at the bottom and then we pipetted 430 microlitres of warm grind buffer to each tube. We placed it in a warm water bath for 10 minutes, shaking it in between and then we pipetted 130 microlitres of potassium acetate into the tubes, mixed them up and placed them in an ice bath. We centrifuged it again for another 20 minutes, labelled a new set of microcentrifuge tubes and pipetted the supernatant into the new tubes, each to its labelled tube. We pipetted 1.5x volume (of the supernatant) of the binding buffer, pipetted all of those into labelled Epoch spin column tubes and centrifuged them. We washed the DNA with ethanol, adding about 500 microlitres of 70% Ethanol to the tubes and centrifuged them, twice to be extra clean (be sure to remove the liquid from the bottom!!). We centrifuged it again, with no liquid this time, to remove any leftover ethanol. We removed the top blue part (the collection tubes) and placed them in clearly labelled microcentrifuge tubes, added 100 microlitres of warm sterile water, centrifuged it again for another 2 minutes to get the elute DNA, which is then placed in ice.

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