Install on Ubuntu 22.04:
python3 -m pip install --user adafruit-ampy How to use an Oxford Nanopore MinION to extract DNA from river water and determine which bacteria live in it Pre-sequencing preparation by 
Ciro Santilli 37 Updated 2025-07-16
Ciro Santilli 37 Updated 2025-07-16One cool thing we did in this procedure was to use magnetic separation with magnetic beads to further concentrate the DNA: Figure 1. "GE MagRack 6 pipetting.".
The beads are coated to stick to the DNA, which allows us to easily extract the DNA from the rest of the solution. This is cool, but bio people are borderline obsessed by those beads! Go figure!
Then we prepared the DNA for sequencing with the Oxford Nanopore specific part: Oxford Nanopore SQK-LSK109 Ligation Sequencing Kit.
How to use an Oxford Nanopore MinION to extract DNA from river water and determine which bacteria live in it PCR verification with gel electrophoresis by Ciro Santilli 37 Updated 2025-07-16
Ciro Santilli 37 Updated 2025-07-16For this reason, it is wise to verify that certain steps are correct whenever possible.
Gel electrophoresis separates molecules by their charge-to-mass ratio. It is one of those ultra common lab procedures!
Since we know that we amplified the 16S regions which we know the rough size of (there might be a bit of variability across species, but not that much), we were expecting to see a big band at that size.
And that is exactly what we saw!
First we had to prepare the gel, put the gel comb, and pipette the samples into wells present in the gel:
To see the DNA, we added ethidium bromide to the samples, which is a substance that that both binds to DNA and is fluorescent.
Because it interacts heavily with DNA, ethidium bromide is a mutagen, and the biology people sure did treat the dedicated electrophoresis bench area with respect! Figure 4. "Gel electrophoresis dedicated bench area to prevent ethidium bromide contamination.".
Gel electrophoresis dedicated bench area to prevent ethidium bromide contamination.
Source. 
Gel electrophoresis dedicated waste bin for centrifuge tubes and pipette tips contaminated with ethidium bromide.
Source. The UV transilluminator we used to shoot UV light into the gel was the Fisher Scientific UVP LM-26E Benchtop 2UV Transilluminator. The fluorescent substance then emitted a light we can see.
As barely seen at Figure 8. "Fischer Scientific UVP LM-26E Benchtop 2UV Transilluminator illuminated gel." due to bad photo quality due to lack of light, there is one strong green line, which compared to the ladder matches our expected 16S length. What we saw it with the naked eyes was very clear however.
How to use an Oxford Nanopore MinION to extract DNA from river water and determine which bacteria live in it Post filtration purification by Ciro Santilli 37 Updated 2025-07-16
Ciro Santilli 37 Updated 2025-07-16After filtration, all DNA should present in the filter, so we cut the paper up with scissors and put the pieces into an Eppendorf: Video 1. "Cutting vacuum pump filter and placing it in Eppendorf".
Cutting vacuum pump filter and placing it in Eppendorf
. Source. 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.
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 1. "White precipitate formed with Qiagen DNeasy PowerWater Kit".
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.
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:
file 5.36 however does use it to display file type as explained at: stackoverflow.com/questions/34519521/why-does-gcc-create-a-shared-object-instead-of-an-executable-binary-according-to/55704865#55704865Please refer to Video "Linus Torvalds saying "Nvidia Fuck You" (2012)".
Does not happen every time, only some times. Can't figure out why. Usually happens when has suspended for a longer time.
bugs.launchpad.net/ubuntu/+source/nvidia-graphics-drivers-470/+bug/1946303 sounds like a likely report, Nvidia driver version 470, but can't find those error messages anywhere. The last line of:once was:which is when sleep starts.
journalctl -o short-precise -k -b -1PM: suspend entry (deep)This suggests that it is not a video bug then, seems that it is not waking up at all? Gotta try to SSH into it. OK. I did SSH into it, and that was fine, so it is just the video that won't start.
PM: suspend exitbugs.launchpad.net/ubuntu/+source/linux/+bug/1949977 is another possible bug, based on kernel version. I'm running 5.13, which is one of the failing versions on the report. Can't find any interesting dmesg though.
In another crash:had the following interesting lines:and there was a corresponding
journalctl -o short-precise -k -b -1nvidia-modeset: WARNING: GPU:0: Lost display notification (0:0x00000000); continuing.
[24307.640014] NVRM: GPU at PCI:0000:01:00: GPU-18af74bb-7c72-ff70-e447-87d48378ea20
[24307.640018] NVRM: Xid (PCI:0000:01:00): 79, pid=8828, GPU has fallen off the bus.
[24307.640021] NVRM: GPU 0000:01:00.0: GPU has fallen off the bus.
[24328.054022] nvidia-modeset: ERROR: GPU:0: The requested configuration of display devices (LGD (DP-4)) is not supported on this GPU.
[repeats several more times]
[24328.056767] nvidia-modeset: ERROR: GPU:0: The requested configuration of display devices (LGD (DP-4)) is not supported on this GPU.
[24328.056951] nvidia-modeset: ERROR: GPU:0: Failed to query display engine channel state: 0x0000927c:0:0:0x0000000f
[24328.056955] nvidia-modeset: ERROR: GPU:0: Failed to query display engine channel state: 0x0000927c:1:0:0x0000000f
[24328.056959] nvidia-modeset: ERROR: GPU:0: Failed to query display engine channel state: 0x0000927c:2:0:0x0000000f
[24328.056962] nvidia-modeset: ERROR: GPU:0: Failed to query display engine channel state: 0x0000927c:3:0:0x0000000f
[24328.056983] nvidia-modeset: ERROR: GPU:0: DP-4: Failed to disable DisplayPort audio stream-0
[24328.056992] nvidia-modeset: ERROR: GPU:0: Failed to query display engine channel state: 0x0000947d:0:0:0x0000000f/var/crash/_usr_sbin_gdm3.0.crash.Related "GPU has fallen off the bus": askubuntu.com/questions/868321/gpu-has-fallen-off-the-bus-nvidia
There are unlisted articles, also show them or only show them.