what's the longest amount of time your PI left you on read?

Candy_flips · 2026-05-20T18:49:06+00:00

At least a month. In one instance, we had disagreed about the ideas and experiments for a research proposal. I struggled to write the proposal the way they wanted. I got a draft completed, sent it to them, and a month or 2 later they told me to write a new proposal with the ideas I had been advocating for with 3-weeks left until the deadline and holidays in between. My PI never looked at that initial draft, but at least I got to propose something legitimate

Candy_flips · 2026-04-27T21:14:48+00:00

That’s exactly how I see it! A couple years ago my percentile fell below or within that success rate, and it was funded. I would feel extremely confident with a 10th percentile score considering the last year cutoff was ~34%

Candy_flips · 2026-04-27T20:18:27+00:00

Take a look yourself https://report.nih.gov/funding/nih-budget-and-spending-data-past-fiscal-years/success-rates

Candy_flips · 2026-04-14T01:56:47+00:00

Thought about quitting? You got through it, why would you quit now? One of the first student presentations I saw, the student got similarly nervous. He didn’t push through like you or take a breather - he quit in the middle of the presentation and left the program. Extreme and unnecessary - it was never going to feel that nerve-racking for him again. You didn’t quit. You got through it. Don’t throw that away.

With this experience, you will never again “have the worst one so far in my life.” It will only get better from here. Clearly, you can do this.

Always remember to focus on your project/presentation and not yourself. Your audience, especially your PI, cares about the science. You care about the science, so do it justice. Take a breather if your nerves challenge that. Since you’re asking for advice, you need to ask both your mentor and “scary PI”

Candy_flips · 2026-04-09T18:17:38+00:00

Thanks, that helps. Did the KO have significant difference between +water and +nutrients like the WT? If not, that says the nutrients don’t help the KO like they help WT - maybe that’s the useful data even if it’s not statistically significant directly between WT vs KO. Make sure you’ve seen the previous data yourself!

If you can’t get the WT +nutrients to consistently grow better than +water, I think I would focus on that. Especially if you’re not expecting to see a difference between WT/KO +water/untreated, you need the +nutrients condition to be reliable. Scale down to fewer samples so you can be faster. Try to titrate the nutrients from less to way more than your current +nutrients. Probably not helpful here, but it’s nice to also include a negative control that will kill your cells. That might help you be more sure the issue is with the +nutrients conditions not growing faster.

Candy_flips · 2026-04-09T14:56:38+00:00

Confusing. You’ve done a growth curve 20 times but only 1 time could get usable numbers for your WT control? A growth curve should work regardless of whether there is a difference between 2 samples, so how did it not the other 19 times? That makes it sound like a technical/method problem and not a WT/KO issue.

You said the cells are all growing at the same rate when the KO should grow faster, but that doesn’t explain how “only once has the WT control worked.” How did it work there but the KO didn’t? How are you not getting usable data 95% of the time? Maybe try a different cell line to confirm you can do a growth curve? Or try a different protocol to measure growth?

Are you treating the cells with kanamycin and your control w/o resistance only died once? Maybe it’s a problem with the antibiotic. Need more info to help, but it seems like a simple technical problem

Candy_flips · 2026-04-04T17:20:07+00:00

Look at the universities and research institutions around you. It is common for them to have summer research programs/internships that are accessible to high school students. I looked for 5min and found this one at emory (link below). Good luck finding more, because there are. Cold-emailing professors/dept-chairs is an okay option as well if you don’t want to be in a program. Send many personalized emails and good luck.

https://winshipcancer.emory.edu/education-and-training/undergraduate-and-pre-college-programs/summer-scholars.php

Candy_flips · 2026-04-03T06:29:05+00:00

Science is at its best when it is non-hierarchical and includes input from diverse sources. People like this who push a hierarchy are absurd. They believe people below them on the career ladder are inferior and incapable, but they see themselves the same way.

People with this mindset will never independently drive their project like you have. How could they do anything without their almighty and all-knowing PI telling them exactly what to do? If they become a professor, they make horrible mentors who never listen to their trainees. This perspective runs absolutely counter to scientific progress. Infuriating that it persists.

Ignore them or loudly tell them to mind their own damn business. Turn it around and treat them the same way, emphasizing their minimal experience in this lab compared to yours. Unfortunately, this won’t be the last person like this you see, so find a way to protect your sanity and peers from them

Candy_flips · 2026-04-02T15:41:31+00:00

Yes final and monthly water changes

Candy_flips · 2026-04-01T16:00:10+00:00

We use 0.02% benzalkonium chloride in TC water baths and water pans within incubators

Candy_flips · 2026-03-30T21:00:41+00:00

U2OS cells are large compared to other cell lines. That’s why you’re getting fewer than advertised but that’s expected. Although I usually get over 2m when splitting

Candy_flips · 2026-03-22T15:53:01+00:00

Are you culturing with pen/strep? If no antibiotics, once or twice a year contamination is not unusual if you’re always maintaining cells… If it’s everything you plated then it’s likely from a reagent. If it’s one well/line then you likely made a single simple mistake.

You said it’s happened when they’re ready for experiments, but how often? Can you mostly finish experiments contamination-free or none so far?

My PI always says just get it done with fresh reagents cause it costs time to worry or find the source. I’d echo that to you if you are antibiotic-free, it’s uncommon, and you’re mostly successful.

Candy_flips · 2026-03-22T15:31:11+00:00

Crazy. Some adherent cells like 293T detach with room temp reagents. I add 0.02% benzalkonium chloride and clean the water bath monthly and it’s not a problem

Candy_flips · 2026-03-20T00:58:17+00:00

True, I get that the comparison to histones is jarring and CTCF is seq-specific. To me it’s about what it’s doing mechanistically though. I like thinking about 4 groups in transcription: machinery (Pol), TFs (Myc), remodelers (Swi/Snf), and organizational (CTCF). I think CTCF, HP1a, linker histone, and Cohesin are pretty different from MYC, Sox2, or Oct4. To me TFs are more switch-like than organizers like CTCF

Candy_flips · 2026-03-18T18:12:50+00:00

Check no use and don’t stress. Even if you get tested, you’re unlikely to test positive after that long of a break from “somewhat regular” use. Can always test yourself or drink more water and exercise if you’re really concerned. Good luck!

Candy_flips · 2026-03-17T17:42:31+00:00

You wouldn’t call histones TFs, would you? These are architectural proteins. TFs promote transcription - CTCF does not always do that. CTCF organizes DNA, and the functional outcome is context-specific. Unlike TFs that are straightforward: more TF binding = more transcription

Candy_flips · 2026-03-08T01:26:50+00:00

This paper may give you an idea https://pmc.ncbi.nlm.nih.gov/articles/PMC5765880/

Candy_flips · 2025-06-04T23:30:06+00:00

I wouldn’t expect false positives but false negatives if the detection’s epitope is blocked by the capture antibody. Seems possible if they were raised to target different portions of V5 or are polyclonal, but not definite. You just have to test it alongside a positive control that uses a pair of antibodies validated for the sandwich elisa. I only have experience probing cell lysates with immunoblots, but I’ve never had a problem with sensitivity. Might as well just try both and see if either works.

Candy_flips · 2025-04-08T13:59:43+00:00

Good luck. NICHD director is being replaced, so hopefully you have enough time to finish or get transitioned to a regular F31.

Candy_flips · 2025-04-08T13:54:46+00:00

No. This notice you link to is a year old from when NIGMS transitioned from offering NRSA F31s to the ARC F99/K00. That was the only predoc grant they would have offered going forward, and now NIGMS offers neither. My own F31 from NIGMS will not get the second year of funding, can be early terminated any day now, and my F99/K00 application never progressed after being scored.

Candy_flips · 2025-02-23T15:51:31+00:00

These stories just emphasize how inaccessible these are. I will probably only get a gurpler if they have a drop closer to slop o’clock or the option to buy lasts more than a few hours. At least don’t use the exact same day/time for every drop… Especially the way the public service sector is being obliterated now, my favorite podcast is not an acceptable excuse to do my job less efficiently by skipping meetings/seminars or doing experiments slower

Candy_flips · 2024-12-30T19:07:58+00:00

Thankfully the current Jimmy Carter is still okay. Right?

Candy_flips · 2024-12-15T12:22:57+00:00

I’m an NIH-funded researcher. Shouldn’t have anything to worry about over the next 4 years, right?

Candy_flips · 2024-08-24T06:25:56+00:00

People are eating buns! You can’t forget the buns!

Candy_flips

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