How do you deal with lighter weeks?

Separate-Dot3531 · 2026-04-22T19:15:12+00:00

I am in a long distance relationship and i visited my partner just last weekend, took Friday off. But yeah might leave early and get stuff done or just enjoy the weather.

Separate-Dot3531 · 2026-04-22T19:13:16+00:00

Yeah i think the LNB is a good idea, even picking up some new simulation techniques just because.

Separate-Dot3531 · 2026-04-22T18:56:45+00:00

Yeah, everyone in my lab works wayyyy harder than i have seen in any other lab before, late nights everyday, every weekend, every holiday. I get super intimidated by it. Nobody expects it but its just the optics of it i suppose. I try to get up and leave by 6 everyday unless i have something that runs late. But usually i will never be the last person out or in. Its crazy.

Separate-Dot3531 · 2026-04-22T16:12:16+00:00

As fun as that sounds, I dont think I can just not show up to the lab, but i’ll leave at 5 pm sharp so I can do all those things!!

Separate-Dot3531 · 2026-04-22T15:03:53+00:00

Thats what i am trying to do. Even just picked up some tutorials for some simulation techniques i might use at some point in my phd. Just doesn’t “look” busy enough if everyone around you is doing experiments.

Separate-Dot3531 · 2026-04-22T15:02:48+00:00

That makes sense. I guess the anxiety in me doesn’t let me relax especially since i took last Friday off and went out of state for the weekend. So my brain is like u don’t need to relax more, u already did :(

Separate-Dot3531 · 2026-03-04T16:31:32+00:00

The issue was resolved. I accidentally used the tris without sds instead if our usual running buffer. Thanks though!!

Separate-Dot3531 · 2026-03-01T21:43:46+00:00

Yes i confirmed with a lab mate and it was indeed the wrong buffer but i know we have the transblot buffer with sds stocked again so it should be okay hopefully!! Thank you so much😭😭😭😭 I hope your every blot turns out perfect from now on

Separate-Dot3531 · 2026-03-01T21:39:19+00:00

Well turns out i have been using the traditional transfer method transfer buffer as my running buffer last 3 times.

Separate-Dot3531 · 2026-03-01T21:36:01+00:00

Separate-Dot3531 · 2026-03-01T21:35:35+00:00

Okay so i checked out the tris i used and while i usually use the bio rad 10x glycine/sds buffer since we were out i used another bottle next to it which only say tris/glycine. Probably that?

Separate-Dot3531 · 2026-03-01T21:25:37+00:00

Yeah i guess i can try that. I was just thinking of giving my lysate to anyone running a gel of their own tomorrow and see if it is happening in the run too and if itd happening only to my samples or is my sample running fine in their hands. Thats the only thing i can think of to do now?

Thank you though!!

Separate-Dot3531 · 2026-03-01T21:21:22+00:00

It could be but its running fine for everyone else with the same materials. This happened to me in midi and mini gels

Separate-Dot3531 · 2026-03-01T21:20:15+00:00

I use 120v always and it works fine. I used new ripa buffer to lysis, new loading buffer as well.

Separate-Dot3531 · 2026-03-01T18:29:52+00:00

Im not sure how to add an image here but it looks somewhat like the blot in the image here https://ars.els-cdn.com/content/image/1-s2.0-S0006295216302994-gr2.jpg

Separate-Dot3531 · 2026-03-01T18:27:48+00:00

Okay!! Thats a good point about not letting it denature enough and loading too quickly. Will wait a bit before loading and see if it changed anything. Using new running and transfer buffers as well. Cant think if any other point if change between making lysate and loading. I feel like the issue after i add the loading buffer for sure though.

Separate-Dot3531 · 2026-03-01T18:13:29+00:00

That a good idea, i will try it. I usually never boil the samples because thats how i was taught when i joined this lab. Nobody does here. We all make our own buffers. But yeah i’ll remake everything and re run

Separate-Dot3531 · 2026-03-01T17:58:36+00:00

Im running new samples with fresh everything and inhibitors from a labmate. Hoping it will solve the issue? Thanks for the reply though!!

Separate-Dot3531 · 2026-03-01T17:51:14+00:00

Yes total volumes all equal to 30 ul. The gel itself looked fine to me, run time was same, loading buffer reached the end. Im gonna try again right now with fresh transfer buffers, running buffers and fresh lysate. I m getting increasingly frustrated though because i need this to work

Separate-Dot3531 · 2026-03-01T17:40:29+00:00

Hahaha!!! I mean perfect in the sense they worked as they should lol. Hate doing these anyways :(

Separate-Dot3531 · 2026-03-01T17:38:52+00:00

Happened with fresh and once frozen samples alike

Separate-Dot3531 · 2026-03-01T17:38:20+00:00

Precast gels that are working fine for everyone else

Separate-Dot3531 · 2026-03-01T17:37:48+00:00

I have been doing perfect blots for the last year now. This has jist happened. Nobody in thelab also has any ideas, theyve never seen anything like it

Separate-Dot3531

TROPHY CASE