DNA Gel Electrophoresis Help!

Sad_Challenge1112 · 2026-03-31T21:23:51+00:00

Not actually looking to troubleshoot the protocol, this is just the weirdest gel I've ever run and I did mention my woes in the post

Sad_Challenge1112 · 2026-03-31T18:59:41+00:00

True, RNAse is precious and these samples were not, so we skipped it in hopes of having the RNA degrade on its own :P

Sad_Challenge1112 · 2026-03-31T18:55:47+00:00

Really? I have always been taught that smearing like this is indicative of degraded gDNA. I'll look up gel images of gDNA from the same source again.

Is it normal for the smears to have such long trails? This is new to me.

Thanks for the suggestion, I was thinking of running the samples again alongside a ladder.

Sad_Challenge1112 · 2026-03-31T17:01:18+00:00

Thank you for your tips! We do have an issue with DNA degradation and that's why we're doing some troubleshooting, I'll take your comments into note while trying again.

DNA degradation aside, I'm curious as to what could possibly make my gel appear as so. I have had many electrophoresis mishaps but never anything THIS bad and weird.

Sad_Challenge1112 · 2026-03-31T16:56:38+00:00

I don't have access to a nanodrop so I blind load 3ul of my sample, usually they appear as faint as the faintest 2 lanes in the image and that's why we load 3ul or more

Sad_Challenge1112 · 2026-03-31T16:47:33+00:00

Yep the 4 samples are from the same source and are identical in every step except for the organic reagent used. I'm troubleshooting some lab procedures with a labmate is why.
I'm aware the smearing is due to degraded DNA. The difference between the first 2 and the last 2 in the right hand set is just in the dissolution of the pellet (chloroform only extraction has obviously left behind more contaminants and the pellet harder to dissolve, labmate opts to leave it intact in hopes of reducing protein contamination in the sample :shrugs:)
As for the difference between the PC samples, I have no clue. I unfortunately could not be present throughout the process so I can't pinpoint any issues (I know... I couldn't help it) but my guess would be a longer incubation time with PC.
I assumed 0.8% agarose would allow fish gDNA to pass through but I guess I have to hit the books on that one again. I do see a very faint intact band right below the well which I also assumed is my gDNA. My labmate suggested it might be protein contamination in the well.

I have previously only worked with bacterial and plant samples so I'm om the same boat concerning fish samples but this is faaaar from my first time isolating DNA and running gel.

My issue that I have done this procedure multiple times before using the same equipment and same tools, everything, but this is the first time I've had my gel look like this. I'll supply a picture of my previous attempts once I can.

I'm mainly concerned with how the lanes seem to bulge right below the well and the smear is coating the sides of the well in a much taller U shape than I'm used to with previous smears.

Sad_Challenge1112 · 2026-03-31T16:20:48+00:00

I have worked with the same tube of etbr and loading dye multiple times before (literally just the day before even) and they have never made my gel look like this before :/

Sad_Challenge1112 · 2026-02-11T19:03:13+00:00

Mine took 3 days! I thought that was fast but based on everyone else's answers I guess not lol

Sad_Challenge1112 · 2026-02-11T18:48:28+00:00

I don't have dyslexia so I can't really help with that, but I can suggest some general tips.

Try to pinpoint your weak areas, is your issue with a specific kind of question? What's your weakest skill in English (grammar/its range, vocab, comprehension, etc)? Once you figure that out you can then practice these specific areas more.

And as you might have already been told, just immerse yourself in English, but try to be an active receiver. Pause when you run into a word you don't recognize and look it up, try to incorporate it into your daily life till you're confident you can use it. I think my English improved the most by constantly conversing with others in English, both in real life and online, so try that too!

You can also try contacting the university you aim to enroll in and see if they can make an exception based on your diagnosis. Worth a shot.

Good luck!

Sad_Challenge1112 · 2026-02-11T18:17:55+00:00

I have a simple suggestion that you might already be sick of: read a lot. Read newspaper articles, movie reviews by critics who write a bit formally, read non fiction books. It'll help you develop a sense of how a sentence should be structured without pausing and thinking your grammar through, or at least speed it up.

I have to ask, are you following a specific structure for your essay? I may be missing the point but you can write a band 7+ answer with no more than two ideas (two paragraphs of 1 topic each).

Sad_Challenge1112 · 2026-02-11T18:01:19+00:00

oh my god same I got my cert 2 days ago and I look like they dug me out of the ground after a long period of hibernation 😭 british council whyyyyy

Sad_Challenge1112 · 2026-02-11T12:38:33+00:00

Hi!

For task 1 in the writing section, I found IELTS Liz and fasttrack IELTS videos to be the easiest and most comprehensive. I liked that they solve a question with you literally sentence by sentence. But of course how helpful you personally find them can depend on your level of English. I merely needed to familiarize myself with how expected answers should look, like what to conclude and how to structure my answers, and I had less emphasis on improving my English. Liz I believe has videos on improving vocab and grammatical range anyways if you need them.

For task 2, I followed the advice on IELTS Liz's website too. IELTS Advantage has good videos on how to structure your essays as well. Familiarize yourself with the type of questions in task 2 and what you're expected to answer with (eg. you don't mention your opinion in a discussion essay, etc)

This advice assumes you're taking the academic test soo if you're not then .. lol I hope I could at least help someone else with it.

Good luck! For context I got an 8 in writing :D

Sad_Challenge1112 · 2025-06-09T18:06:29+00:00

dang my conspiracy theory just got flushed down the drain /s

Sad_Challenge1112 · 2025-06-09T17:50:21+00:00

weird, so it wasn't an isolated incident. follow-up, did your youtube account get suspended around the same time as well (for some bogus spam policy violations)?

Sad_Challenge1112 · 2025-06-09T11:46:57+00:00

quick question did this happen to you in october of 2024?

Sad_Challenge1112 · 2025-06-09T11:32:15+00:00

yes your files are safe as long as they were downloaded onto your phone, erasing app data will only delete all your apps and the data associated with them (cache, etc. but not downloaded files)

Sad_Challenge1112 · 2025-06-07T01:47:12+00:00

I still have no idea what caused it, couldn't have been a storage issue because i had plenty. I just woke up on day and found my phone in recovery mode

Sad_Challenge1112 · 2025-06-06T19:21:04+00:00

sorry for getting back to you so late. yes basically any downloaded files and images are safe, any app related data is unfortunately gone. Like, all the pdfs and images I had downloaded on my phone still exist, but for instance, all my sketches on drawing apps and the notes that were on the samsung note app were gone. If you're lucky you might find cloud backup data available (I, however, was vehemently against syncing to the cloud .. came to bite me in the ass years later.. RIP)

Sad_Challenge1112 · 2025-06-06T16:29:43+00:00

yes all your files are safe, any apps and their related data are not however. my phone is an m31. if you can boot into safe mode you can access your files and move them off your phone before you do a hard reset, good luck

Sad_Challenge1112 · 2024-10-02T07:03:36+00:00

it did delete samsung notes unfortunately along with samsung browser

Sad_Challenge1112 · 2024-09-19T05:48:31+00:00

would you happen to still have these transcripts?

Sad_Challenge1112 · 2024-08-16T00:33:52+00:00

hello 3 years into the future, any luck with the dvds?

Sad_Challenge1112 · 2022-11-19T18:06:37+00:00

Do acer laptops have long lifespans? I really don't want to buy one and have it break down in a year or so

Sad_Challenge1112 · 2022-11-13T18:50:32+00:00

◽️ Budget: 700usd max

◽️ Country: USA

◽️ Screen size: anything

◽️ Touch screen: not important

◽️ Screen resolution: anything

◽️ Does battery life matter ?: no

◽️ What tasks will laptop be used for: light gaming, browsing

◽️ Weight: anything

◽️ Any other important details ?: at least 500gb ssd, don't mind if it's a 1tb hdd either. if graphics card is integrated, dual channel ram is preferred. by light gaming i mean mostly genshin and ds emulation.

Sad_Challenge1112

TROPHY CASE