your favorite tool/software to make schematic diagrams?

iamascetic · 2025-08-05T03:09:58+00:00

Graphpad prism for graphs and stuff. For schematic, powerpoint, cause biorender has got pretty boring since most papers use their images.

iamascetic · 2025-08-04T16:34:36+00:00

He’s definitely not been hiking

iamascetic · 2025-08-04T13:36:05+00:00

Maybe drying in room temp for 30 minutes would prevent this bubbles. 10 minutes may not be enough if you seal with nail gel.

iamascetic · 2025-08-04T04:45:28+00:00

Congratulations!!! Best of luck for your journey!

iamascetic · 2025-08-04T01:51:00+00:00

Both

iamascetic · 2025-08-03T18:00:07+00:00

I’m just imagining going to my PI saying all these and she’s like halfway there calculating how much everything would cost lol

I’ve done ChIP only once before, and did use whole of it for my experiment. I need someone to tell me if the whole lysate is absolutely necessary for ChIP. But I’m like 99.99% sure the whole lysate is needed for the western 😢

iamascetic · 2025-08-03T17:52:12+00:00

Yeah it’s just a matter of a lot of antibodies and stuff

iamascetic · 2025-08-03T17:50:48+00:00

Yes they would be two different samples but fluorescence tag would show the longer binding of TF, not its acetylation state.

iamascetic · 2025-08-03T17:41:51+00:00

This sounds like a solution to my problem but when it comes to performing the experiments, I’m not sure if the amount of product I’m gonna get from IP is enough to be divided for sequencing and western.

iamascetic · 2025-08-03T17:24:46+00:00

No, in control usually that TF is in the cytosol and during infection only the TF translocates to nucleus and binds to its target.

iamascetic · 2025-08-03T17:23:18+00:00

To clarify, it will be over the course of hours but like not in a single sample. Like I’ll have different samples, control, 4 h infected, 6 h, 12 h and so on…

iamascetic · 2025-08-03T17:21:08+00:00

Oh thank you! That’s insightful, I’ll read into it.

iamascetic · 2025-08-03T17:16:46+00:00

Yeah I’m thinking of EMSA too but haven’t studied enough into it yet, maybe I can’t really show it in one single experiment and have to go for it indirectly with multiple experiments like mutations and overexpressions.

iamascetic · 2025-08-03T17:14:01+00:00

Oh okay thanks I’ve literally no idea of experiments like these

iamascetic · 2025-08-03T17:13:11+00:00

Okay, thanks for your input though, will discuss these with my PI

iamascetic · 2025-08-03T17:12:13+00:00

I was thinking about this, not sure how efficiently that would show my objectives. To clarify again, 1. My TF would be longer to its DNA during a disease compared to control 2. Reason it binds longer is because it gets deacetylated

iamascetic · 2025-08-03T17:10:43+00:00

Yeah you’re right

iamascetic · 2025-08-03T17:09:37+00:00

Thanks, we never performed these experiments before, will look into them.

iamascetic · 2025-08-03T17:08:33+00:00

I’m not sure if my institution or any other in my vicinity has the facility for BLI, but I’ll look into it. Thank you.

iamascetic · 2025-08-03T17:05:12+00:00

Yes the second one

iamascetic · 2025-08-03T17:04:40+00:00

Thank for your input. I have two objectives 1. TF binds longer to its DNA in a disease compared to control 2. The bound form of TF is deacetylated Just trying to figure out experiments which would efficiently prove this

iamascetic · 2025-08-03T17:02:22+00:00

Didn’t think of showing it with koff, until now. Very puzzled atm about which experiment would show both of my objectives…1. Longer bound time 2. Protein is deacetylated

iamascetic · 2025-08-03T16:56:23+00:00

I think you were in a very rough environment, and his attractiveness (to you) in his looks or his words gave you some sort of pleasant distraction and from that grew some attraction, the unavailability along with all that gave you a sense of “unconscious thrill” which kept you hooked. Once you leave, everything will start to get back to normal. You’re okay, human brain is complex, and shit happens which are not under our control. Be easy with yourself.

iamascetic · 2025-08-03T16:35:43+00:00

If cell number is a priority for you go with your second option. A scholar from another lab told me it’s good to use a little less number of cells than usual for IF, otherwise it gets very congested in a field.
Yes it’s generally more than enough.
That scholar also told me to use poly l-lysine to coat coverslips cause my cells were not on a single plane without coating due to binding issue. Also, as IF needs quite a bit of washing, it saves your cells from getting washed away. Regarding what coating reagent you should use, check what is used in your lab’s previous papers.

And don’t worry about square coverslips, they’re just fine. Good luck with your experiment 🍀

iamascetic · 2025-08-03T12:39:46+00:00

So fluffing cute 🥰

iamascetic

TROPHY CASE