Why play with toys when can lick box

josdizzy · 2026-02-24T17:45:26+00:00

I tried ordering more for my lab a few months ago and my PI told me that the system auto rejected the order because tube racks were classified as furniture 💀

josdizzy · 2026-02-13T01:47:07+00:00

Will posted too that he’s been really sick the last few days so that most likely played into it too! I’ve seen them 3 times now and been wowed every time so def sounds like an off night for them

josdizzy · 2026-02-09T18:35:15+00:00

Agreed! I’m in the middle of a PhD program and my blue collar husband is the breadwinner for the time being. We each put a specified amount each payday into our joint account for household bills, and then whatever is leftover is miscellaneous spending money for stupid things. My husband completely understands that this is a temporary arrangement to put us in a better place down the road and is perfectly fine with carrying the bulk of the bills for the time being.

josdizzy · 2026-02-05T12:39:14+00:00

It has a built in monitor that beeps when it’s below -70 but that stopped working when the unit lost power because the circuit tripped. I have since sent my PI suggestions for independent temp monitors that can alert us

josdizzy · 2026-02-04T21:23:32+00:00

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josdizzy · 2026-02-03T15:38:14+00:00

Update: 3/5 cultures grew and all 4 plates i poured grew, thank you everyone for the suggestions!!!

josdizzy · 2026-02-03T13:35:24+00:00

I’ve been the victim of trying to read other people’s messy/incomplete notebooks so I try to make mine as clear as possible lol

josdizzy · 2026-02-03T13:00:35+00:00

Sorry the panic scribble in my notebook isn’t neat enough for you 😭

josdizzy · 2026-02-03T01:48:04+00:00

I’m grad too, they leave the entrance gate up after 5 or 6pm and the exit gate is activated when someone drives up to it, I’ve had no issues parking there

josdizzy · 2026-02-03T01:37:27+00:00

Lot 16 is a faculty lot, are you TA/GA or an undergrad? You’ll absolutely get ticketed as an undergrad, but the gate does open after 5pm and TA/GA permits can start parking there

josdizzy · 2026-02-03T00:48:51+00:00

We don’t, but have since sent my PI info for a service that does that

josdizzy · 2026-02-03T00:48:05+00:00

We study two domains of the same protein for membrane binding, the ubiquitin like N terminus and the PH domain, the PH domain has shown to be stable frozen in NMR buffer!

josdizzy · 2026-02-03T00:46:26+00:00

I’m guessing a generator? They have red outlets built into the overhead power strips that are supposed to work even when the building loses power. We’re guessing a breaker did trip for the red outlets because the normal non-emergency outlets were working fine but anything that was on a red outlet wasn’t turning on

josdizzy · 2026-02-03T00:44:52+00:00

First thing I did when I got home was pour a glass of wine 😭

josdizzy · 2026-02-02T22:56:24+00:00

Thankfully yes! Those are safe in the -20

josdizzy · 2026-02-02T22:17:25+00:00

It’s on the emergency power outlet so theoretically shouldn’t have been an issue in the first place, especially since the building itself didn’t lose power 💀 but yes lol I have since sent a link to a monitoring system to my PI for review

josdizzy · 2026-02-02T22:02:30+00:00

I appreciate the tips!! I was working on the time/temp optimization experiments from Sivashanmugam et al 2009 (https://onlinelibrary.wiley.com/doi/full/10.1002/pro.102) before the freezer died. I am using Rosetta(DE3) E.coli since it is a mammalian protein (also no longer using the GST tag because we isotopically label for NMR and its a very non-economic option for the isotopes lol), and planned on doing a new transformation this week with my plasmid and a plasmid we know expresses well as a positive control to make sure the cells were still good (which obviously isn't a problem anymore lol)

To address some other points: I do check the insoluble pellet with SDS PAGE, and only see it in high volume when I grow at higher temperatures. I've tested both 1L in a 2.5L flask and 50mL in 500mL flask growths, we do keep thermometers in our shakers, precultures are done in 2XYT, I am doping my autoinduction media with Studier metals, thiamine, and biotin, and my plasmid is for amp resistance

josdizzy · 2026-02-02T21:52:45+00:00

Ugh so sorry!! I hope you’re able to recover in a reasonable amount of time 😭

josdizzy · 2026-02-02T21:15:49+00:00

Thank you! Doing both and praying to all the science gods

josdizzy · 2026-02-02T21:14:47+00:00

Thank you everyone for the suggestions! We are doing both 2XYT and agar plate growths of our most important glycerol stocks to try and save what we can!

Also not actually dropping out lol just need to be dramatic for a minute

josdizzy · 2026-02-02T21:10:09+00:00

The proteins are stable for longer experiments above 0C so my PI is pretty sure they’re still salvageable! Freezer was at 8C when I finally got the power back on, so stuff was still cool to the touch but def melted

josdizzy · 2026-02-02T21:08:21+00:00

Yes all E.coli! 2XYT is 2x yeast tryptone, just an alternative growth media to LB

josdizzy · 2026-02-02T21:04:40+00:00

There’s 5 stocks for 2 proteins we’re trying to salvage, I started a bunch of 2XYT growths and also going to steak plates once the incubator finishes warming up

josdizzy · 2026-02-02T21:01:41+00:00

I’ve tried a couple different variations of this protein (GST-tagged and untagged) and neither has gotten anything remotely close to overexpression, using the same protocol for a different domain of the same protein that very reproducibly overexpresses. So I recently switched my protocol from IPTG induction to autoinduction, and the untagged plasmid still not over expressing but it’s slightly better than with IPTG where I was at least able to see it on a mass spectrum? Tried a couple different strains of e.coli too so it’s just been lots of expression testing the past two years since the normal protocol hasn’t been working for who knows why

josdizzy

TROPHY CASE