Experiments on the hierarchical separation of mitochondria
Experiments on the hierarchical separation of mitochondria
Fractionation (Fractionation) is a gradual sedimentation by centrifugation from low to high speed. First use low speed to make the larger particles precipitate, and then use a higher speed, will float in the supernatant particles precipitate down, so that a variety of cellular structures, such as the nucleus, mitochondria, etc. can be separated.
Operation method
high speed centrifugation
Materials and Instruments
mitochondria Move 1. Remove the high-speed centrifuge tube containing the supernatant from the beaker with ice, level it, and centrifuge it at 17,000 rpm for 20 minutes, discard the supernatant, and retain the precipitate. For more product details, please visit Aladdin Scientific website.
centrifuge
2. Add 1 ml of 0.25 mol/L sucrose-0.003 mol/L calcium chloride solution, blow the suspension with a pipette and centrifuge at 17,000 rpm for 20 minutes.
3. aspirate the supernatant into another test tube, retain the precipitate, and add 0.1 ml of 0.25 mol/L sucrose-0.003 mol/L calcium chloride solution and mix to form a suspension (toothpick may be used).
4. Take the supernatant and precipitate suspension, put one drop each on a clean slide (labeled ① and ② smears, respectively), and cover the slide with one drop each of 0.02% Janus Green B stain for 20 minutes.