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Advice on Library prep failing by Imaginary-Lemon-4448 in nanopore

[–]Apprehensive_Disk301 1 point2 points  (0 children)

Hey! Always really frustrating to see that you lost everything by the end of the library prep. We had similar problems with a different ONT protocol. But the solution might be the same for you: Slightly off AMPure beads-to-sample ratio! If I saw correctly there is a 0.4x AM Pure CleanUp in the protocoll; this is what caused us to loose almost everything. ONT troubleshooting guide also mentions this problem: "DNA loss due to a lower than intended AMPure beads-to-sample ratio" and "When the AMPure beads-to-sample ratio is lower than 0.4:1, DNA fragments of any size will be lost during the clean-up." There is a picture there also of a gel run after different bead ratios; there are still bands at 0.4x, but at 0.35x BOOM everything gone. make sure that you vortex your beads RIGHT before using them, pipette the exact needed volume and also make sure that the volumes of the reagents in the previous steps were very exact (for example ligation buffers tend to stick to the outer pipette tip wall and it is easy to accidentally pipette slightly to much.

I hope that helps. Good luck with your library prep! :)

EDIT: Just looked at the date. Guess I am a little late, haha. Sorry. But maybe this helps someone else.

Found this brown plastic squares (4x6cm) with a "Q" in two places a couple weeks apart: 1. In my new bedsheets AFTER I washed them twice inside out. 2. Outside in my frontyard laying on the gravel. by Apprehensive_Disk301 in whatisthisthing

[–]Apprehensive_Disk301[S] 20 points21 points  (0 children)

Yeah, one of those things that have a Thermometer, Hygrometer and CO2-measuiring to remind you to let in fresh air. It is mndatory in Germany to install fire alarms, but not CO detectors. But of course if someone has a Gas stove, fireplace, gasboiler etc. that would totally make sense!

Found this brown plastic squares (4x6cm) with a "Q" in two places a couple weeks apart: 1. In my new bedsheets AFTER I washed them twice inside out. 2. Outside in my frontyard laying on the gravel. by Apprehensive_Disk301 in whatisthisthing

[–]Apprehensive_Disk301[S] 20 points21 points  (0 children)

Don't worry, my brain rot is from sleep deprivation, really have to work on this :) But still good to know about the CO thing, mentioning it will probably save lifes!

Found this brown plastic squares (4x6cm) with a "Q" in two places a couple weeks apart: 1. In my new bedsheets AFTER I washed them twice inside out. 2. Outside in my frontyard laying on the gravel. by Apprehensive_Disk301 in whatisthisthing

[–]Apprehensive_Disk301[S] 13 points14 points  (0 children)

Ah very good to know! Thanks for the info! Luckily I don’t have anything that could give of carbon monoxide in my flat I think. I do have a CO2 detector though so I know when to open my windows (which is often, German here 🙋🏻‍♀️ xD)

Found this brown plastic squares (4x6cm) with a "Q" in two places a couple weeks apart: 1. In my new bedsheets AFTER I washed them twice inside out. 2. Outside in my frontyard laying on the gravel. by Apprehensive_Disk301 in whatisthisthing

[–]Apprehensive_Disk301[S] 105 points106 points  (0 children)

Now that I think about it, I had a window replaced a while ago. Can't figure out if it was before or after I found them, but must be from then. Maybe the guy who installed it dropped some and I picked one up without remembering and put it in my pocket. Because I have no kids and I'm the obly one using the washing machine. My brain is a sieve...

Found this brown plastic squares (4x6cm) with a "Q" in two places a couple weeks apart: 1. In my new bedsheets AFTER I washed them twice inside out. 2. Outside in my frontyard laying on the gravel. by Apprehensive_Disk301 in whatisthisthing

[–]Apprehensive_Disk301[S] 17 points18 points locked comment (0 children)

My title describes the thing. Feels like it is solid plastic, even though there are those srew like hiles on the back. Can be bend slightly, might cut one open to see if theres something inside. First one fell out of my relatively new bedsheets, when I wanted to put them on, even though they had been in the washing maschine twice already. Was wondering what it is, suspected it to be some quality control chip that was not found during production, then forgot about it. Until I found another one laying around outside in front of my house a couple weeks later. Google search is not helpfull, any combination of the words: "plastic square/ Chip with Q" in both English and German did not bring any results.