Ciro Santilli  Sponsor 中国独裁统治 China Dictatorship 新疆改造中心、六四事件、法轮功、郝海东、709大抓捕、2015巴拿马文件 邓家贵、低端人口、西藏骚乱
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. "Cutting vacuum pump filter and placing it in Eppendorf".
Video 3.
Cutting vacuum pump filter and placing it in Eppendorf
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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.
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 cell and free the DNA.
Video 4. Source.
Video 5. Source.
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.
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. "White precipitate formed with Qiagen DNeasy PowerWater Kit".
Figure 17.
White precipitate formed with Qiagen DNeasy PowerWater Kit
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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.
In our case, we used a VWR Micro Star 17 microcentrifuge for those steps:
Figure 18.
VWR Micro Star 17 microcentrifuge.
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Figure 19.
VWR Micro Star 17 microcentrifuge loading.
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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:
Figure 20.
Biochrom SimpliNano spectrophotometer loading sample.
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Figure 21.
Biochrom SimpliNano spectrophotometer result readout.
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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:
Figure 22.
Minus 20 fridge storing samples.
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