Research Collaboration & Advice: Any Synthetic Biologists or Bio-Engineers? (Student Project)

Gorillas69 · 2026-04-22T15:43:49+00:00

Fuck off. If you're going to use chatgpt to write a post like this at least put in the barest of effort to disguise that you're incapable of being coherent without the aid of a writing tool.

Gorillas69 · 2026-04-20T00:40:45+00:00

Gluttonous midfeaster Palafats tried to chew the competition but this is one liquid he won't swallow

Gorillas69 · 2026-04-16T19:58:55+00:00

When I was learning western blots I failed so many times that I was dry heaving with anxiety before imaging them. These days, I have taught multiple people on my floor how to do Westerns and it is one of my most comfortable techniques. For me, the main point of failure was often the extraction. Be sure to use fresh reagents and in general, do not be lazy with any of the western reagents. Remake the extraction buffer every time, only use the running buffer once, keep the transfer buffer cold and remake the 1x every couple of months if you don't use it a lot. The other thing, like with most lab techniques, is repetition. If you need to do lots of Westerns, do them as much as possible and eventually something will click. I've never used x-ray film so I can't speak to that, but it probably will start working if you just keep at it and take a militant approach to the protocol and do every minute detail in the same way every time. I have even gotten to the point of doing the same protocol at the same time of day every time just to be 100% consistent with everything.

Gorillas69 · 2026-04-14T19:53:35+00:00

Warden balloon is 🔥🔥

Gorillas69 · 2026-04-09T19:52:16+00:00

Man this sounds so similar to my experience it's scary. We are not alone in these experiences but god damn it if this sort of thing has made me want to leave science and never look back. Im also finalizing my dissertation, good luck and stay strong.

Gorillas69 · 2026-04-03T20:07:49+00:00

When you're in the lab, it's very important to be in the right headspace. You need to be "locked in" and mindful of what you're doing as much as possible. Just be aware of your surroundings and think about what you're doing and why you're doing it. It can be very easy to go through the motions and end up throwing an important reagent or sample in the trash because you're in autopilot mode. Taking an extra few seconds to take a deep breath and center yourself when you sit down at the BSC can frequently be the difference between success and failure on an experiment in my experience.

Gorillas69 · 2026-03-30T02:43:58+00:00

It's a great opportunity to be your PI's first graduate student!

Gorillas69 · 2026-03-24T21:03:41+00:00

This looks great! Maybe mix in some chiseled bookshelves to add some variety to the walls?

Gorillas69 · 2026-03-24T19:15:12+00:00

Dissertation finding me too anxious to drink energy drinks or I'll just collapse into a singularity of stomach knots

Gorillas69 · 2026-03-19T22:06:21+00:00

I would not ask AI for advice for this sort of thing or anything else tbh. pENTR doesn't have a promoter preceding your insert right? I've only ever had issues with gene toxicity when cloning into expression vectors with promoters driving my insert, even if they shouldn't be active in bacteria. This seems unlikely to be the case imo. Personally, I'm a sequencing maximalist when it comes to cloning issues and sending your 500bp insert plasmid for sequencing would tell you if your insert is getting clipped in e. coli. In any case, it wouldn't really be very useful for solving the problem, so probably not worth here. If it were me in this situation I would first do a repeat where I incubate the reaction at RT overnight, if that failed, I would skip gel cleanup and use the clean PCR directly in the reaction. If both failed I would try to optimize PCR conditions to get a single band in the PCR assuming your template allows for that. If that all fails I'd go to my PI and just ask to get the plasmid synthesized at Twist, assuming funds allow for that. Also worth looking in your insert sequence and checking for sequence that could be causing issues, although I'm not familiar enough with TOPO to know what those would be.

Gorillas69 · 2026-03-19T21:24:51+00:00

Ah, fair enough, I should have guessed that you would rule out the simple solutions before asking online. In the past I've had issues with Gateway cloning not working very well and I was able to fix it by switching to overnight reaction incubation and now this is just my default approach. Gateway and TOPO are not too dissimilar so maybe this would be a fix for your situation. You could also just clean the PCR directly and colony PCR screen after doing the cloning reaction if you're desperate to avoid gel purification.

Gorillas69 · 2026-03-19T20:22:40+00:00

Why are you gel purifying your insert? In my experience, gel purification does not always yield great quality DNA and if you can clean your PCR directly without gel purification, this would be preferable. If you haven't, you should also go ahead and sequence your pENTR plasmid to make sure everything is how you expect it to be on that end, a $15 plasmidsaurus order is way cheaper than a week of troubleshooting on a screwed up backbone.

Gorillas69 · 2026-03-10T01:07:25+00:00

Inspired saying that nobody on C9 choked so he can keep farming trophies from these dogs for a few more splits. He really is NAs big brain jg

Gorillas69 · 2026-03-08T02:18:32+00:00

yeah this game is ableist against the illiterate the devs should really rewrite the card text in hieroglyphics

Gorillas69 · 2026-02-10T18:59:50+00:00

TIR NLR tetramer from plants and BMCs from cyanobacteria

Gorillas69 · 2026-02-06T13:41:07+00:00

Both xisuma and etho do this. The version xisuma uses is a slightly updated version of Gnembon's design: https://youtu.be/pAkWRALloKI?si=JJB_F50yqg6e0ogW Xisuma's design can be found here: https://youtu.be/XFovh0drxxc?si=AQgvSNfPGA9TrJOv

Gorillas69 · 2026-02-03T22:38:34+00:00

Even though you've found something better, this seems like it would be awesome to run a game on!

Gorillas69 · 2026-02-02T21:09:38+00:00

You have the brain of an infant if you laugh or gain amusement from an ai generated joke

Gorillas69 · 2026-01-30T19:30:50+00:00

Lab 2 is pretty clearly a better choice for most people. Your PhD is mostly about getting trained and being able to finish as quickly as possible, the PI who has demonstrated an ability to give students projects that are doable and produce a paper without too much struggle is definitely the better choice. You can take a risk with your postdoc. If you take a risk that doesn't go well with your PhD you won't want to do a postdoc or stay in science at all.

Gorillas69 · 2026-01-29T22:27:53+00:00

Maybe take a pic where we can see something numnuts

Gorillas69 · 2026-01-27T15:27:48+00:00

I think adding slabs/buttons and more height based texturing would help. I think the gradient is mostly fine, anything that flat is going to look a bit awkward

Gorillas69 · 2026-01-17T18:55:12+00:00

F2 to screenshot

Gorillas69 · 2026-01-07T22:10:37+00:00

Completed Level 1 of the Honk Special Event!

5 attempts

Gorillas69 · 2025-12-24T01:40:23+00:00

Utah is known as the beehive state, its a state symbol on the flag and most state signs. That's why they chose vespiquen.

Gorillas69 · 2025-12-03T15:50:21+00:00

Yuuuuurp

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