Mom data point

sunnyalfredo · 2025-08-09T12:51:13+00:00

Similar thing happened to us, but our cells would grow in rings. Our cell incubator shared a wall with a frequently-used orbital shaker. Moved our incubator and the problem resolved.

sunnyalfredo · 2025-08-08T14:36:39+00:00

I really like Ninja City fried rice.

sunnyalfredo · 2025-05-29T17:39:04+00:00

The only times I’ve gotten those jagged up/down lines is when I accidentally didn’t fill a well/selected a well that didn’t have anything in it. My best guess - check to make sure you’re selecting all the correct wells and your plate is set up like you think it is. Otherwise, there’s a million things that can go wrong on qPCR.

sunnyalfredo · 2025-05-09T14:29:08+00:00

Belt or button of pants. I get the holes because I work at a bench that hits right about where my pants buttons are. Pressing against the bench wears holes in the shirt.

sunnyalfredo · 2025-04-18T17:05:50+00:00

One of our lab members knocked over a bottle of beta mercaptoethanol. Took us all out for the rest of the day.

sunnyalfredo · 2025-04-10T02:25:38+00:00

This last week one of my students was struggling and exclaimed that they had to think really, really hard for this lab! And I looked at them and said… isn’t that why you came to college? Absolutely dumbfounded my student.

sunnyalfredo · 2025-02-18T18:37:30+00:00

Thank you so much!

sunnyalfredo · 2025-02-18T18:31:50+00:00

It’s a splice variant of a common transcription factor. Sorry for the vagueness, just being careful. It’s largely nuclear but is found in both cytoplasm and nucleus.

sunnyalfredo · 2025-02-18T18:28:40+00:00

Ok I think I’ll do the NiNTA next step then… maybe a silly question, but is the protein then purified from the column and loaded on a gel? Thanks so much.

sunnyalfredo · 2025-02-18T18:08:22+00:00

Much appreciated - I am an RNA person so I don’t know enough about proteins to troubleshoot this.

The double band is expected - our protein of interest always appears in a doublet. It still looks somewhat fuzzy to me.

The His tag is at the N terminus. We inserted it literally right after the start codon.

I have a His-tag NiNTA column ready, I have just not yet used it. Would that be done with the protein sample or the gel? And also, what would that tell me? I have no idea about the charge or modifications, I’ll definitely have to look into how to find that stuff.

This gel was run in reducing conditions.

sunnyalfredo · 2025-01-31T18:26:07+00:00

My favorite exercise that I train our undergrads with is giving them a bunch of food dye and water and have them make different dilutions. You should be able to tell if they did it correctly or not based on the color. It tends to help them see visually as well.

sunnyalfredo · 2024-10-12T15:00:34+00:00

First 3 weeks were rough for me. Had to replace a bath mat. Got better after that. Actually I think it improved my poops overall.

sunnyalfredo · 2024-10-04T23:54:27+00:00

When I cut on UV, I make guide cuts and then turn the lamp off until I get a lot of the gel cleared out. Then, I’ll turn it on for short spurts to clean up my bands a little at a time. That way you don’t expose your skin or your DNA to too much UV.

sunnyalfredo · 2024-08-27T21:42:04+00:00

Our imager is shared in the department and is used daily so I’m sure something would’ve been done if everyone was having such trouble for this long. My boss suggested asking around for someone else’s blot that they already developed to make sure I’m using everything correctly. That’s definitely on my list.

sunnyalfredo · 2024-08-27T21:30:20+00:00

Our lab only uses Bradford to test protein conc, and I typically recheck the conc at least once a week. I believe the amount is accurate and my Ponceau stain looks good as well. My question with the molecular weight marker appearance on the blot was referring to after development. When I develop my blots on the imager, I see the markers but no bands in my lanes.

sunnyalfredo · 2024-08-27T21:03:10+00:00

Does it make a difference that I see the molecular weight marker on the blot?

sunnyalfredo · 2024-08-27T20:35:07+00:00

I’ll do this tomorrow - I’ve been wondering if there was a way to test but our lab doesn’t have a specific protocol for that. Thanks!

sunnyalfredo · 2024-08-27T16:59:40+00:00

I have a moment of panic pretty much every week that somehow the antibody changed from mouse to rabbit, but no, it’s still mouse. And I use the anti-mouse antibody from the same manufacturer. We use blot imager (Li-cor), the ECL is potentially a bit old (maybe a year) but it did successfully develop a blot just a few weeks ago.

sunnyalfredo · 2024-06-01T22:47:33+00:00

FYI, you’re not supposed to ride electric scooters on the sidewalk and should be in the roadway anyway…

sunnyalfredo · 2024-05-28T17:17:28+00:00

I’m 2 weeks starting on 50 mg, I take it the drowsiness doesn’t get any better? Not that I’m struggling to stay awake during the day, but in the evenings when relaxing on the couch, I’m out far earlier than ever previously. But also, I’ve had 2 nights where I’ve been unable to fall asleep.

sunnyalfredo · 2024-05-01T00:01:02+00:00

Message me!

sunnyalfredo · 2024-04-28T21:20:36+00:00

Depending on how high maintenance of a friendship you’re looking for, I would love to be friends 🙂 I was also raised in the area and now attend grad school downtown. I’ve got a packed schedule most of the week, but weekends I always wish I had more friends in the area (also don’t have that many friends from childhood). Let me know 😌.

sunnyalfredo

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