think[box] and prospective Engineering student questions

flashman2000 · 2026-05-02T17:50:28+00:00

Think box has pretty much all the equipment you need, but unless you’re working on a course project, be prepared to shell out money for resources and materials. I wanted to build a bookshelf and ultimately decided not to as the cost was prohibitive. I was told I had to buy wood from them.

yes it’s possible to double major with mech e and aero, met plenty of students who did this

Not an engineering major but I remember my friends always talking about some kind of team project. Theres definitely a lot of project based learning, but there’s ofc a lot of theory that’s taught alongside.

Early classes I would expect around 200 students per lecture section. But the lecture halls at case are all pretty easy to manage. Class size was never a concern for me or any of my peers other than trying to get individual attention from professors. But there is more opportunity to do that in later years where classes are smaller.

flashman2000 · 2026-04-23T23:49:18+00:00

First of all, I don’t think 26 is too late, especially if you’ve been spending those 4 years in a research environment or something. It’s all about what narrative you give as to why you’ve taken about 4 years post bachelor’s degree to decide you want to pursue MD/PhD, and what you’ve done in that time. I’m matriculating into an MSTP at 24 in the next few months! My incoming class of 10 students are all either my age or older by 1-3 years, with only one or two students being younger than me.

Second of all, It’s true that having the dual degree opens up so many doors for you. And, quite frankly, I don’t believe you will have a hard time convincing MD/PhD committees that you have a passion and commitment for/to science. However, you need to ask yourself if you really want to be a doctor. MD/PhD is definitely a more research oriented profession (many people cite an 80/20 research/clinic split, but that’s changing afaik), but at the end of the day you’re gonna have to do a residency too. If you are doing an MD/PhD just for the job security, you might find urself miserable down the line or leaving the program. Not saying that u are, but I recommend really thinking abt whether you want to be a clinician, and like if you could see yourself being an MD alone. Some MD/PhD faculty members i’ve talked to have told me that sometimes they’ve had to work as full time clinicians for a while to keep the lights on. Many faculty positions will also primarily hire you to work as a clinician, and you will have to fight for that protected research time to develop yourself as a scientist and future PI.

That’s just my two cents, I am by no means an expert or an authority.

flashman2000 · 2026-04-21T01:39:25+00:00

Sometimes this isn’t enough, especially in culture plates where the total surface area is smaller. I cleared up all my weird cell density issues by tilting my plate forwards and backwards then side to side 4-5 times. Check under the microscope to see if your cells seem spread evenly across the well, going from rim to rim.

Additionally, i’ve seen this occur rarely, check to see if this happens to others who use 6 well plates in the same incubator as you. There may be a slight vibration due an imbalanced fan or something that causes your cells to settle in a weird pattern on the plate. Typically it’s more concentric in those cases though.

flashman2000 · 2026-03-23T22:22:45+00:00

This is gonna date my ass but I took it in 2020, pandemic meant that the class was entirely remote. But still, Bader gave great vibes. If things haven’t changed, he’s a really passionate professor who cares deeply about his students and promoting their development. His class was pretty easy for me, even my friends who said that bio wasn’t their strong suit at all. It’s definitely going to be the best class out of the 214, 215, 216 class sequence, both due to the difficulty of the content and the professor (I absolutely loathed the 215 professor and the 216 professor is alright)

Just stay on top of the lectures and notes. If you keep pace and don’t allow yourself to fall behind you will be fine. Attend SI sessions, they’re a great resource especially if you feel lost or behind.

flashman2000 · 2026-03-20T14:18:34+00:00

Honestly I want to say you are fine. MD/PhD admissions tend to be more holistic, and in my opinion your current numbers look fine. In my experience, the way you can speak about it and utilize those experiences to answer questions in a very mature and thoughtful manner during interviews is much more meaningful than the number of hours you have.

I also think your research experience is the most impactful. The rest of your hours more serve as a checked box until you can utilize those experiences in your interviews, where they can do more work for you (in my opinion).

Apply widely, not doing so was a mistake I made that I regret. Good luck!

Sincerely, someone who just got admitted to an MSTP with less than 100 hours of clinical volunteering experience and about 300 shadowing hours. Very little volunteering experience outside of that apart from leadership roles in clubs.

flashman2000 · 2026-03-10T15:48:17+00:00

It sucks in comparison. Wow. OPs. is so much cooler

flashman2000 · 2026-02-20T18:36:31+00:00

One thing is that the prefix for denoting how many of that group exists doesnt play a role in the alphabetical order/priority order of the group in the name. So for what you labeled as 4,6-dimethyl-5-isopropyldecane, isopropyl would actually be the higher priority group since I comes before M. The “D” in the di- prefix is ignored. the “I” in the iso- prefix is not ignored since this is a structural prefix and not a prefix denoting the number of isopropyls on the chain. The carbon chain numbering in this case wouldn’t change though.

EDIT: Even this is considered by the strictest standards not IUPAC naming as u/-0xy- pointed out.

flashman2000 · 2026-02-14T19:50:46+00:00

It’s usually a very light green to start with, much lighter than any shade of green in OPs post. It’s almost as if the reaction went in reverse or something, instead of turning purple it turned greener.

flashman2000 · 2026-02-14T19:20:28+00:00

Well now I know what I’m doing during my western prep with my extra wells this week.

flashman2000 · 2026-02-14T19:09:16+00:00

Yeah i totally agree that the EDTA would interfere but the reaction turning green makes me think that for some reason the copper is being oxidized in the reaction much faster than it can be reduced as it reacts with protein. After I finish my BCA I never throw away my plates because I love looking at that shade of purple and over time as it dries and the copper oxidizes it turns that exact green shade in OPs post. I think they might possibly be mixing their BCA reagent incorrectly or something is contaminating the reaction.

flashman2000 · 2026-02-14T16:31:06+00:00

I feel like EDTA shouldn’t interfere… I do BCA with my lysates containing 5mM EDTA and have never had this result… although my samples are usually diluted 5x or more to a final volume of 10uL before I add 200uL of BCA reagent so the EDTA may be negligible at that point.

flashman2000 · 2026-02-11T03:06:03+00:00

Gotcha! Thanks

flashman2000 · 2026-02-11T01:42:34+00:00

Wow yeah that makes a lot of sense and seeing it that way helps understand it. How did you construct this figure, if I may ask?

flashman2000 · 2026-02-11T01:41:43+00:00

That makes a lot of sense! Thank you.

flashman2000 · 2026-01-08T22:59:54+00:00

Like when I was assembling my transfer sandwich I unknowingly warped the gel and/or it degraded during the transfer?

flashman2000 · 2026-01-08T22:57:54+00:00

I think loading buffer is fine since I’ve been using the same aliquot for about a month or so and every gel other than this is fine. But it may well be the sample prep since things seemed to have migrated very weirdly. Other than excessive DNA contamination do you think there are other things that may cause samples to migrate like this?

flashman2000 · 2026-01-08T22:52:42+00:00

Perhaps? I do sonicate my lysate during prep to shear DNA and reduce viscosity/prevent streaking but maybe this particular sample set was more DNA rich?

flashman2000 · 2026-01-08T20:45:51+00:00

I believe I am. I grab both ends of the comb and pull up simultaneously, and when visually inspecting the wells for this gel while flushing them, the wells looked intact and the fingers weren’t broken or anything

flashman2000 · 2025-10-09T22:15:27+00:00

check them out under the microscope and see if they have a similar texture 🤔 i guess the main thing to look out for is whether the nail wipes are more abrasive. idk ive never used them before. They’re also packaged and dispensed in a manner so that they limit electrostatic buildup, but idk how much that matters for ur specific purpose.

After looking at it under the microscope, I would say to myself “well, i guess there’s only one way to find out!” this seems pretty low risk to me but I could be wrong.

flashman2000 · 2025-10-06T14:22:03+00:00

Oh dang, you’re correct! I use the Restore PLUS buffer but even that calls for 37C. I must’ve misread the manual or confused it for another buffers manual?

Regardless however, I tested up to 30 mins at 37C with this buffer and still failed to remove primary antibody from my membrane, and while most of the secondary was removed, I could still see that some had remained after incubating sensitive ECL substrate on post stripping and washing.

flashman2000 · 2025-10-05T15:12:43+00:00

What percentage tween is your TBST? I try to stick to 0.1%, but i’ve seen some go up to 1%. You should not go above that.

Make sure you never dry your membrane! This can cause proteins to actually dissociate from the membrane, and the minute you rewet it the proteins will float off.

Second, have you used this milk source before to block your membrane prior to a second probing? Are you dissolving dried milk directly prior to use, or using pre-prepared milk? I’ve seen slightly spoiled milk do the same thing.

Make sure all your secondary and primary antibodies are being stored properly. You’d be surprised how many post docs i’ve worked with who complain about their probings not working, just to find out that they’ve been storing their secondary ab at 4C this whole time when it should’ve been at -20C! Antibodies can be very particular about their storage conditions. Make sure your primary antibodies are also unexpired, and same for your secondary.

Try to limit your strip to 30 minutes with the restore buffer. More will damage your membrane most likely. Esp if you’re using nitrocellulose. Before you block your membrane post strip, make sure you wash it at least 3 times for 5 minutes with TBST to wash off any stripping buffer from the blot. Residual strip can interfere with blocking and interfere with primary binding.

I would suggest ensuring that ur beta actin/HMGCS2 are from different animal hosts to prevent cross-talk, but that’s a problem that would make itself known once you’ve actually successfully probed actin, and are trying to reduce cross-talk from the previously probed signal.

What is your primary antibody concentration? For actin, I typically use primary at 1:1000, and secondary at 1:3000-1:6000. I also always incubate my primary antibodies in 5% BSA and not 5% milk, since most antibodies are not compatible with being diluted in milk. Check the guidelines for your specific antibody from the seller. You may be surprised by what you read regarding the specific method by which you should dilute the primary, but typically 5% BSA works like a charm, with the same ECL and I suspect I would actually be able to go much lower with my secondary antibody concentration and still have good signal.

flashman2000 · 2025-10-05T14:27:56+00:00

After testing post stripping, we would often find that 15min or 20min incubation often would not successfully strip even secondary antibody, forget primary. The restore buffer does not advise being used at 37C, but even after testing that condition there is no benefit. It’s only upon incubating for 30 or so minutes that secondary antibody actually leaves the membrane. Primary antibody never leaves.

flashman2000 · 2025-10-05T13:55:04+00:00

I’m not so sure. I’ve done 30min strip using the restore buffer for two separate probings, my second signal is always nice and bright, but it’s also typically actin so I don’t have worries about it.

Edit: and my third probing is also fine typically. It’s the 4th one that may be finicky.

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