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Post filtration purification

| 🗖 nosplit | ↑ parent "DNA extraction" | words: 302
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After filtration, all DNA should present in the filter, so we cut the paper up with scissors and put the pieces into an Eppendorf: Video 3. "Vacuum pump filter cut and place in eppendorf.".
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Video 3. Vacuum pump filter cut and place in eppendorf. Source.
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Now that we had the DNA in Eppendorfs, we were ready to continue the purification in a simpler and more standardized lab pipeline fashion.
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First we added some small specialized beads and chemicals to the water and shook them Eppendorfs hard in a Scientific Industries Inc. Vortex-Genie 2 machine to break the cells and free the DNA.
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Video 4. Scientific Industries Inc Vortex-Genie 2 loading. Source.
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Video 5. Scientific Industries Inc Vortex-Genie 2 running. Source.
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Once that was done, we added several reagents which split the solution into two phases: one containing the DNA and the other not. We would then pipette the phase with the DNA out to the next Eppendorf, and continue the process.
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In one step for example, the DNA was present as a white precipitate at the bottom of the tube, and we threw away the supernatant liquid: Figure 17. "Qiagen DNeasy PowerWater Kit White Precipitate.".
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Figure 17. Qiagen DNeasy PowerWater Kit White Precipitate. Source.
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At various stages, centrifuging was also necessary. Much like the previous vacuum pump step, this adds extra gravity to speed up the separation of phases with different molecular masses.
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In our case, we used a VWR Micro Star 17 microcentrifuge for those steps:
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Figure 18. VWR Micro Star 17 microcentrifuge. Source.
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Figure 19. VWR Micro Star 17 microcentrifuge loading. Source.
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Then, when we had finally finished all the purification steps, we measured the quantity of DNA with a Biochrom SimpliNano spectrophotometer to check that the purification went well:
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Figure 20. Biochrom SimpliNano spectrophotometer loading sample. Source.
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Figure 21. Biochrom SimpliNano spectrophotometer result readout. Source.
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And because the readings were good, we put it in our -20 C fridge to preserve it until the second day of the workshop and called it a day:
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Figure 22. Minus 20 fridge storing samples. Source.
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