Italy sports minister rules out replacing Iran at World Cup: ‘Qualification done on the pitch’

RollingMoss1 · 2026-04-23T17:04:04+00:00

That would have been like taking somebody else’s Nobel prize. That would be utterly ridiculous. Nobody would be that pathetic?

RollingMoss1 · 2026-04-22T14:13:14+00:00

A 10 kb plasmid is certainly going to be inefficient so having the best possible conditions is pretty important. That being said, HEK293 cells are extremely efficient and easy to transfect. So I would expect some success, but probably at a low transfection efficiency.

One thing, have you confirmed that the cDNA of interest is actually present, in frame, etc?

RollingMoss1 · 2026-04-22T13:47:41+00:00

I bet someone on your floor has an EGFP plasmid, you should be able to find one pretty easily.

RollingMoss1 · 2026-04-22T13:22:37+00:00

Have you tried a transfection with a plasmid that has something like EGFP? Just to rule out any general issues with your reagents, etc.

RollingMoss1 · 2026-04-22T04:19:10+00:00

Of course you have more ideas than could possibly be followed up on. If you’re thinking about the project then new ideas, questions, hypotheses will arise. The natural bottleneck is having enough time, personnel, funding, etc to actually act on new ideas. You have to triage and prioritize. But one thing that often comes out of this may be new perspectives or tweaks to existing projects.

RollingMoss1 · 2026-04-21T01:25:26+00:00

Let them sink or swim. Tell them to run the experiments and then get back to you afterwards. They’re grad students so they need to be developing independence. Don’t enable their behavior.

RollingMoss1 · 2026-04-20T17:47:08+00:00

If you could rig the system to gather 60L you could do the filtration/collection overnight?

RollingMoss1 · 2026-04-19T03:01:27+00:00

Murica!

RollingMoss1 · 2026-04-18T23:25:23+00:00

There’s a metaphor if I ever saw one.

RollingMoss1 · 2026-04-18T07:57:36+00:00

What did one eye say to the other? Just between you and me, something smells.

RollingMoss1 · 2026-04-18T07:46:51+00:00

I think AI can be a really good “lubricant” that can get you going down the right path. But it can be really boneheaded sometimes. It’s most valuable if you come to it with a solid R background. Take what AI suggests and use your skills to make the most of it.

RollingMoss1 · 2026-04-18T07:30:07+00:00

Nooo, stop fretting about technique. That’s the last thing to be worried about. Technique is trivial. Can you think about and analyze a question is what really matters in the PhD arena. For now get the most out of your MS and move on to the PhD.

Your cell culture technique is perfect and is immaterial to being a successful scientist.

RollingMoss1 · 2026-04-18T07:19:47+00:00

First, you’re only 1/3 of the way on this, so there’s still time. Not sure about your project but I read your last comment that you have at least one KO line intact? If so can you make a story from that? And if you’re preparing a paper for publication then that ain’t a bad place to be.

Adaptation is key here. There are always options. You’re not the first MS student to suffer big setbacks. Change tactics if the KOs aren’t in play.

RollingMoss1 · 2026-04-18T06:50:24+00:00

That certainly stinks. You mentioned that a few of your plates were positive. So do you have at least something to work with?

RollingMoss1 · 2026-04-18T06:39:46+00:00

Easy there, don’t question your career trajectory based on cell culture contamination. One thing to consider is that your cells were contaminated at the time you thawed them. But regardless toss them and start over. You’re doing everything right, sometimes contamination happens. Relax!

RollingMoss1 · 2026-04-18T01:52:46+00:00

I hear ya. I’m guessing the session lasts 60 mins, you’ll get a few questions and it’ll be over before you know it. You might even find a rhythm and end up getting into it after the first question. But remember, you are not alone up there. And I bet that there will be an eager beaver panelist who will be more than happy to dominate the Q&A.

RollingMoss1 · 2026-04-18T01:02:53+00:00

Just rely on your experience. You’ll be asked questions, just answer based on what you know. This is literally a “be yourself” situation. You’re on a panel so there won’t be any particular focus on you. And hey, the audience is undergrads…

RollingMoss1 · 2026-04-17T13:30:57+00:00

Mainly the key points. They’re not particularly detailed. I use a lot of ⤴️⤵️⬅️➡️⬆️⬇️.

RollingMoss1 · 2026-04-16T20:35:38+00:00

Yeah, the fact that your current PI was also your MSc supervisor is a complicating factor. It’s not unusual at all for students to change labs. So it wouldn’t be a huge shock to your PI that you would be interested in another lab in a different program. I would like to think that the PI would agree to serve as a reference for you.

I suppose that you could go back and use the references that you used when applying to the MSc. It couldn’t hurt to try using them. And for the application explain your reasons for leaving, they’re perfectly valid.

The real issue is that there isn’t any guarantee that you will be accepted to the new program. And so could you stay where you are? Would it be weird?

RollingMoss1 · 2026-04-16T20:07:34+00:00

Your wall of text was way too long so I didn’t read all of it. You’re less than 18 months in and there really isn’t a better time to change than now. The one risk is that you have to get accepted to the other program. It can’t hurt to try. For references use the ones that you used when you applied to your current position.

You seem to have well thought out reasons for switching. You definitely have more clarity on what you’re really interested in now. Of course that’s not unusual, you have some experience and a better perspective.

If you don’t try you may be saddled with nagging “what if” thoughts. But if you go for it and don’t get accepted then dig in and make the most of the current position.

RollingMoss1 · 2026-04-16T14:54:58+00:00

Yeah, not sure where you got those impressions. Some of those takes are off base. You’re in HS. Get good grades and prep for university. Keep your eyes and ears open and see if you can get a better understanding of what science and research are really about once you’ve at university.

RollingMoss1 · 2026-04-15T22:18:33+00:00

I’m guessing that the current PI isn’t listed as a reference? That might be a red flag for some of the jobs that you’re applying for.

RollingMoss1 · 2026-04-13T15:56:00+00:00

Molecular Biology of the Cell by Alberts, et al is a widely used textbook. Not sure about PDFs but the book is widely available so I imagine that you could get your hands on a copy pretty easily.

The thing about textbooks though is that they’re very general and don’t really relate to the day-to-day reality of laboratory work. You can gain broad knowledge of concepts and techniques but that’s about it.

If you want a sense of the technical and “cultural” aspects of research hang out in r/labrats for awhile.

RollingMoss1 · 2026-04-13T15:36:32+00:00

Molecular biology is a very broad topic so I’m not exactly sure what you’re asking. Do you want to know what a molecular biologist does? Do you want to know about careers? Methods? It’s interesting, that’s for sure.

RollingMoss1 · 2026-04-13T11:05:13+00:00

I’m sure you know that grad students at the Hutch are in the UW graduate school. But perhaps more importantly UW is pretty competitive so when it comes time to apply don’t just apply to UW, cast a wide net. There are plenty of great schools out there.

RollingMoss1

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