After going through the RNA extraction Trizol protocol, I have 24 samples with a variety of concentrations based on their nanodrop readings ranging from 14ng/up (not so great) to ~300ng/up (much better than I need). My experiments are done in triplicate so I have 8 different RNA sequences and at least 1 out of the 3 for each has a decent concentration so that isn’t my main concern, however, despite having good(ish) concentrations, my 260/280 and 260/230 numbers are much lower than the expected and I have no idea why?

Sample Examples:

Concentration: 147ng/ul 260/280: 1.62 260/230: 1.73

Concentration: 267.2 260/280: 1.79 260/230: 0.93

I’m using the RNA for an RT-PCR and to be honest, I’m going to try it anyway because I have barely any time left to get these experiments done, but it’d be good to know if it’s likely that all hope is lost and it’s going to turn out awfully.

https://reddit.com/link/omdvtw/video/oio836o74ub71/player

One of the most common ways for people to get to know the Nano cryptocurrency is by earning free micro-fractions through faucets. Since Nano is instantaneous, extremely divisible and no fees, this process is totally feasible and inexpensive.

Some months ago I had the need to integrate a nano faucet into a web application. But the current faucets don't provide any kind of API for that and they weren't developed to work well inside iframes either.

That's why I decided to make a new faucet. It is with great pleasure that I present:

https://nanodrop.io

NanoDrop is not only another Nano faucet, but the first Nano Open Source Faucet, transparent and integrable through customizable Checkbox and API.

I used Google's reCaptcha checkbox as inspiration.

Soon anyone will be able to integrate this checkbox on their own website or app! Imagine websites distributing Nano or maybe using this checkbox to sign up users and etc? This is totally possible and extremely beneficial for Nano!

NanoDrop includes:

• Faucet with customizable checkbox.
• QR code reader.
• Nano deposits / donations automatically received.
• Real-time pay table with websockets.
• Anti-spam and anti-bot barriers such as:
  • "Tickets" with amount, ip and timestamp signed using Nano's algorithm
  • Google reCaptcha V2 - forces the user to solve the recaptcha challenge
  • Google reCaptcha V3 - Gives the user a score to ensure they are not a bot
  • Google oAuth - If the user's score is low, it requires login through a Google account
  • Limit per Nano account
  • Limit per IP address
  • Limit per email address
• PoW cache, allowing for always instant transactions.
• Public API for faucet data.

Here is a preview of the process:

https://reddit.com/link/omdvtw/video/swa4m8uf5ub71/player

Of course, a dark theme couldn't be missing either! It automatically adjusts with user settings.

https://preview.redd.it/tot87tjwzub71.png?width=1403&format=png&auto=webp&s=a4356ac713e8f3bfc8994b6b8d04999c2f8bdcc1

Calculation of each drop:

Returns 0.01% of the balance, rounded down.

Example: With a balance of 1.145 Nano, returns 0.0001 instead 0.000145

Or returns the maximum configured amount, by default = 0.01 Nano.

If there is not enough balance, an error is displayed.

[Error Display](https://preview.redd.it/sh2ctmmn5ub71.png?width=346&format=png&amp

So I’m working with blood samples (not human) and trying to get some purified DNA. The blood is frozen and has all the plasma removed so its just a gunky mess in the bottom of the tubes. Total mofo to move into cent tubes. Anyways, on to the mishap. I was running the protocol (regular dna spin prep kit) during the last elution & centrifuge I banged my rack pretty hard on the table. Ended up with some good readings and a few that were really high on the 260/230. Could my hard bang on the spin columns caused some issue with those samples? Im thinking not because it was the last step and everything should have washed out already. I just wanted another opinion. Samples may have just been ass not sure.

Check the new and updated version of the faucet list.

I included a new nano faucet that pays out once!

Does anyone know of a Florida CURA location that has the original NANODROPS in stock- comes in a skinny green dropper bottle. 🙏🏾🙏🏾🙏🏾🙏🏾🙏🏾🙏🏾🙏🏾🙏🏾

I have a NanoDrop 1000 Spectrophotometer that I would like to run on an older machine. I could install Win XP on it, but I 'm concerned with the lack of support. I much rather have it run on Linux, through wine. This way users could still use the browser to access email an other things securely.

Anyone tried this yet?

Just a PSA. If you do a lot of plasmid work and amplification. Visualize your plasmids on a gel to get a more accurate picture of how much quality and concentration of plasmid that you have. Nanodrops will give you a 2x-10x higher estimate than they actually are.

If your lab is rich, invest in a Qubit: that has wonderful accuracy. Otherwise, seriously, start visualizing your plasmids on a gel and you'll start getting incredibly reproducible assays and transfections.

https://www.google.com/amp/s/www.infofueguina.com/salud-bienestar/2019/1/16/un-nuevo-colirio-podria-reemplazar-los-anteojos-segun-investigadores-35886.html Si remodelan la cornea me gustaría pensar que significa también que la aplana, ya que si acaba con el astigmatismo y demás, supongo que también hará lo mismo.. De todos modos les he escrito para saber si ayudaría a pacientes con queratocono, o si están desarrollando algo para el KC, algo que pueda ayudar, de todos modos me interesa mucho y espero que salgan pronto y sean efectivas "Pachymatrix y IVMED-80" pero la maldita burocracia siempre tiene que estar tocando las narices..

If they reshape the cornea I would like to think it also means it flattens it, since if it does away with astigmatism and such, I guess it will also do the same.... Anyway I have written to them to know if it would help patients with keratoconus, or if they are developing something for KC, something that might help, anyway I am very interested and hope they come out soon and are effective "Pachymatrix and IVMED-80" but the damn bureaucracy always has to be touching the noses..

https://mobile.reuters.com/article/amp/idUSKBN28Y0XU?__twitter_impression=true&s=09

Science: where it's okay to screw up as long as you find something useful.

I got a weird result from the NanoDrop for the 260/230 ratio. It was 5,55! For the isolation of plasmid DNA we used the NucleoSpin Plasmid EasyPure. Somehow I'm the only one of my class that had this weird result.

My teacher couldn't really put her finger on it and I can't leave it as it is. Does anyone know how the spectrum of DNA would've looked like if you mixed up the last 2 buffers? Or when you only use one buffer twice and forget the other buffer?

Because a high 260/230 means that there weren't that many organic compounds. The 260/280 ratio was fine, I guess, it was 2,00. But what does the AQ buffer even do then? So many questions..

If anyone has an idea, please let me know! Thank you!

NanoDrop Soectrum (plasmid) DNA from NucleoSpin https://imgur.com/a/XLm0vSP

I picked these up along with the Layer Cake a few days ago, in grapefruit. I started with 10mg just to get a baseline of what to expect. So since nothing at all happened with 10mg when I woke up at 2 am, I took 20mg and I basically lost consciousness lmao!!! Has anyone tried these, and what was your experience like? Since I woke up I backed the dose down to 15mg and I feel like I just vaped a large amount of flower. I feel fantastic! This might outweigh the vile taste of the product! I mean it is seriously foul. But I can overlook it, because I feel like I took a moderate dose of an opioid. I’m rambling because I’m baked so, have a good day all!

Edit: I forgot to add that the effects started to peak at around 15-20 minutes so these really are water soluble and kick in fast unlike edibles. This is just what I’ve been looking for since I am soooo sensitive to edibles.

If yes, does the spectrophotometer account for the pathlength?

Has anyone ever used a nanodrop in the field? Is this an absolute crazy idea or would it be just fine? We were thinking of using a nanodrop lite since its relatively small and doesn't require a computer.

We got a new Nanodrop One, and it's a beautiful machine, but I can not get it connected to our network. According to our IT I need to enable DHCP and set the default gateway, but these settings are not accessible via the interface. The interface is based an Android and fells like a tablet GUI. Doe anybody know a trick how to get into the "real" network settings?

...Asking for a friend...

Hi everyone, today I was using NanoDrop to check DNA conc when I stupidly forgot to close the lid. I was wondering if there were any potential dangers from UV exposure as I didn't close the lid? I feel completely fine with no feeling of burns, but my paranoid self was wanting to make sure.

We typically don't add an RNAse inhibitor to RNA samples in the lab until we make cDNA, but by then the samples have already been on ice for a while for quantifying on Nanodrop.

Would adding an RNAse inhibitor as soon as extraction is finished affect the Nanodrop's readings even if protein is on a different wavelength?

Does anybody here add RNAse inhibitors right after extraction?

I use a nanodrop 1000 in lab however because the software only runs on old windows (7 or vista), I would have to have a 2nd pc to just use the nanodrop which was somewhat annoying. I know, not a huge problem but enough for me to spend a handful of hours searching and trying different solutions. I did come across a few posts here on reddit, however, they were of no help.

The most competent suggestion was to run the software on a VM machine. I tried that but the crux of the issue seemed to be that win10 couldnt recognize and communicate with the machine (connected via USB) so I would not get the option to run the usb connection into the VM machine. I am no computer expert so maybe a PC person could have gotten that to work.

However, I did find a solution that works and am hugely grateful to have found that article. They found a way to make it so that the PC recognizes the USB connection and the software is able to connect to the device without any weird hacks or Virtual machines.

I am here basically to let people know in case there are any OCD SOBs out there that cant get over not being to use the software on a win10 machine.

Here is the link.

https://genomicist.blogspot.com/2019/11/how-to-install-nanodrop-1000-software.html

I wonder how long it will take to measure 200 samples in singlet on a Nanodrop? Anyone know?

Should I be worried about high 260/230 readings for DNA? Mainly in the range of 2.5-3.5. My 260/280 readings are between 1.8-1.9. I know that low 260/230 readings indicated salt and/or phenol, so what would high readings indicate? Protein contamination?

You know what I’m talking about.

Don’t get me wrong, the 2000c is awesome and it saves me boatloads of time.

But still...

Hi everyone! I wasn't sure where to post this but thought /r/biology would be a good place to start.

I'm a grad student doing research in a Genetics-based Cancer (Leukemia) lab and we're having some trouble with our RNA readings.

My labmate was having some inconsistencies in her qPCR readings, so she began a process of elimination to try to figure out what may have caused that. One of the things she is doing is re-measuring the concentration of her RNA samples with the Nanodrop.

Upon re-measuring, she's getting much different readings than what she had when she measured them a year ago but what's even weirder is that she measures each sample exactly the same way multiple times and (most of the time) each reading is quite different. For one sample, she had a very low number, an extremely low negative number, a very high number, and again a low number. The values for absorbances at 260 and 280 seem to be proportionally lower than what she got the first time she measured samples a year ago because the values for 260/280 are mostly around 1.9 which is what she was getting a year ago.

This tells me that the problem cannot be her sample of RNA. The only other factors are the Nanodrop machine itself, and the DEPC water the RNA was suspended in. One other note is that we tried another Nanodrop machine and in general she was having the same problem though not the same exact numbers. We tried to have a tech in our dept. recalibrate the machines but for some reason the machine is not allowing that. My labmate is going to call the company to try to figure out how to calibrate/fix the machine but in the meantime I thought I'd post on here and try my luck.

Has anyone ever had anything similar to this happen to them? What could possibly cause this? Could DEPC water cause this and how would it do so?

Any information at all would be greatly appreciated (READ: I will absorb it like a sponge). I hope I've explained everything clearly. If not, please let me know and I will clarify.

Thanks so much everyone!

EDIT: Thanks again everyone! We talked to our PI today and she played around with the Nanodrop and the samples. A few of you were right in that her concentration of RNA was too high for the Nanodrop and that we should try diluting it out. That worked like a charm and gave us pretty consistent values so her samples are fine but she's going to have to re-do the reverse transcriptase and qPCR reactions.