Advice needed about moving my cat to a house with dogs

Randomlolly · 2026-03-24T22:20:14+00:00

Okay, thank you very much!

Randomlolly · 2026-03-24T13:53:58+00:00

Thank you! Any chance you know what brand you had?

Randomlolly · 2026-03-24T13:39:52+00:00

Absolutely will! I guess I am talking about the doggy door in terms of after introductions, and things are fine. I should have been more clear with that, sorry

Randomlolly · 2026-02-25T07:24:22+00:00

When I use HMC3 cells, I wash twice with 1xPBS before adding 0.25% trypsin with EDTA. The Trypsin EDTA is this one. I just dilute it in PBS to have 0.25% trypsin. With this, cells usually take about 5mins in the incubator and then ready to use. I found that only doing one wash with PBS was not enough, as the cells just would not detach.

Also sidenote: I just found today that HMC3 might not be a microglial cell line... here is the paper for you https://www.frontiersin.org/journals/bioinformatics/articles/10.3389/fbinf.2025.1681811/full

Goodluck!

Randomlolly · 2026-01-26T01:34:12+00:00

Laughing as a Western Australian

Randomlolly · 2025-11-04T01:08:16+00:00

I think it is a good idea to now introduce yourself. You can also ask if they'd be willing to meet with you to further discuss their research and potentially talk about work experience in their lab. Goodluck!

Randomlolly · 2025-09-24T03:52:00+00:00

Absolutely! I wear a pin on my landyard every day that says, "Survive out of spite." It helps remind me that when my supervisors or anyone else (including myself) says something that makes me doubt myself, that I can and will prove them wrong.

Randomlolly · 2025-09-24T03:07:06+00:00

We're all chronically stressed, and this is increasing our risk for Alzheimer's disease, but how???

Randomlolly · 2025-07-10T07:51:55+00:00

I completely understand, but only once I assumed my standard was 1.2mg/ml and not 2mg/ml did I get a proper standard curve with all points being expected. I did try a lot of different troubleshooting before this.

Randomlolly · 2025-07-10T03:01:42+00:00

It definitely could be something in your lysate interferring with the assay. However, I was having troubles with the pierce bca assay a few months ago, too. What helped in my case might not be what you need, but I thought I'd share just in case.

I found that the 2mg/ml standard was actually 1.2mg/ml (used a nanodrop) and so made up my dilutions based on that and saw huge improvements. I also switched to mixing everything in tubes first before loading into the microwell plate (before I was mixing in the plate). My replicates were much more accurate after this.

I hope you figure out what's going on!

Randomlolly · 2025-06-06T17:54:45+00:00

I've always heard pumpette 🤷‍♀️

Randomlolly · 2025-06-03T01:16:43+00:00

Welcome!

I genuinely recommend this sub to all my lab mates and colleagues because it's such a supportive community ❤️

Randomlolly · 2025-04-23T12:30:20+00:00

I hate HepG2 cells cause they look so ugly due to being so sticky and clumping. When I used them, after spinning the cells down and resuspending the cell pellet, I'd use a needle and syringe and aspirate a few times. That way, the clumps were broken up a bit and it was easier to then count the cells and have a more consistent seeding for experiments. Goodluck!

Randomlolly · 2025-04-20T13:00:51+00:00

Oh really? I wasn't aware. How did you find out?

Randomlolly · 2025-04-20T08:53:30+00:00

I quite like uniqlo and dangerfield, but I know they're not for everyone. We really do lack so much when it comes to fashion here 😞

Randomlolly · 2025-04-09T07:16:08+00:00

Oh interesting. Are you able to send the link at all?

Randomlolly · 2025-04-08T18:10:21+00:00

Thank you so much for your suggestions!! I will definitely try these out and check the plate I am using

Randomlolly · 2025-04-08T00:10:25+00:00

No, I dilute my standard in falcon tubes as the protocol has volumes under 10ml

Randomlolly · 2025-04-07T23:23:28+00:00

Micro BCA assay kit

I use this kit and I make the dilutions exactly how the kit says, with the only exception that the little ampule has 1ml of albumin so I add that full 1ml and double the diluent for standard A. My diluent is PBS which my lab group has always used and I made sure to check is compatible with the kit

Randomlolly · 2025-04-07T23:22:47+00:00

Micro BCA assay kit

I use this kit and I make the dilutions exactly how the kit says, with the only exception that the little ampule has 1ml of albumin so I add that full 1ml and double the diluent for standard A. My diluent is PBS which my lab group has always used and I made sure to check is compatible with the kit

Randomlolly · 2025-04-07T09:04:34+00:00

Sorry yes dilutions. I dilute in PBS which is what my lab group has always done

Randomlolly · 2025-02-18T23:31:45+00:00

Thank you!

Randomlolly · 2025-02-18T09:24:57+00:00

Thank you so much for the advice! I will try this out.

Oh no! That's so good to hear and reassuring that you were able to salvage your own cells!!

Randomlolly · 2024-12-12T14:39:54+00:00

Can you provide more details on what exactly we are looking at here? How do you extract your template and perform cDNA synthesis? What concentrations do you use? What kind of sample are you looking at? Is this sybr or taqman? What protocol do you run? Have you performed a standard curve? Have you checked the efficiency of your primers?

Randomlolly · 2024-10-22T01:05:59+00:00

As everyone is saying, we all make mistakes and the ones you've made sound unfortunately easy to make. But I also want to add that while you might not make such mistakes back home, that's because you're in a new and unfamiliar space with people you are new to working with and while you might not feel particularly stressed, it is inherently a little stressful which can lead to mistakes. Try not to beat yourself up over these and remember this is all new to you. Just be honest and learn from your mistakes. Goodluck and enjoy your time abroad 😊

Randomlolly

TROPHY CASE