BIOL 2070 Lecture Notes - Lecture 4: Elution, Microbiological Culture, Reagent
30 Jan 2018
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Pellets were formed from 1 - 5 ml of bacterial culture overnight by centrifugation at. >8000 (6800 x g) rpm for 3 minutes at room temperature (15-25 c). The pelleted bacterial cells were resuspended in 250 l buffer p1 and transferred to a microcentrifuge tube. The solution was mixed with 250 l of buffer p2 by inverting the tube 4-6 times until it became clear. It was ensured that a lysis reaction did not occur for more than 5 minutes. If lyseblue reagent was used, the solution turned blue. After adding 350 l of buffer n3 the solution was mixed immediately and thoroughly by inverting the tube 4-6 times. If lyseblue reagent was used the solution turned colourless. The tube was centrifuged for 10 minutes at 13,000 rpm (~17,900 x g) in a table-top microcentrifuge. By pipetting, 800 l of the supernatant was added to the qiaprep 2. 0 spin column.
