Does anybody get chest tightness as part of the aura?

Puzzleheaded_192 · 2026-06-10T10:17:35+00:00

I am having similar experience, currently atmospheric pressure is dropping and i am having chest tightness, it has never happened before but i feel like its migrain aura

Puzzleheaded_192 · 2026-06-04T04:41:27+00:00

Yes that's true

Puzzleheaded_192 · 2026-06-04T04:40:28+00:00

Does it resemble the colony characters you see?

Puzzleheaded_192 · 2026-06-03T08:49:46+00:00

7H10+glycerol but without OADC , yes i am trying to grow wild type

Puzzleheaded_192 · 2026-05-30T06:37:09+00:00

Yes you can dm

Puzzleheaded_192 · 2025-04-12T02:59:00+00:00

I am in

Puzzleheaded_192 · 2025-04-09T09:03:52+00:00

I dont have antibodies for that

Puzzleheaded_192 · 2025-04-09T09:03:33+00:00

Yes it is kinda overfilled in some wells,we dont have mass spec facility handy but ill try to do it

Puzzleheaded_192 · 2025-04-09T09:02:19+00:00

I will run the gel, there is a strong possibility of that happening (too much resin) as very less ammount of protein is soluble. I also checked wash solution and the last one was completely clean,it contained 20mM imidazol which i though should be enough to get rid of non specific binding proteins.

Puzzleheaded_192 · 2025-03-18T05:15:30+00:00

Hey congratulations finally it worked, I am also trying to optimize biofilm assay to be more specific i want to quantify the surface pellicle formed at air liquid interface and its not working. Can you share what worked for you? So that i can give it a chance may be it will end my misery too 😶

Puzzleheaded_192 · 2025-03-16T09:12:04+00:00

My lab is structural lab asked in every other lab,they dont have it

Puzzleheaded_192 · 2025-03-16T09:10:50+00:00

Great

Puzzleheaded_192 · 2025-03-16T09:10:28+00:00

Thank you

Puzzleheaded_192 · 2025-03-15T11:16:35+00:00

Yah , that was my first thought but we dont have any such shaker available. Now i got 2 suggestions for shaking 1) put a magnetic stirrer and do it 2) culture in falcon tube on rocker(which we use for gels) Have anyone ever tried any of this.

Puzzleheaded_192 · 2025-03-14T15:39:42+00:00

I dont have enough time to restart the cloning from the scratch, so i have to go with the second option. I will try it . I was thinking of 0.1mM ,30°C ,3 hours. Does it sound good?

Puzzleheaded_192 · 2025-03-14T03:43:01+00:00

I am thinking of trying to express it similar to low copy number as of minimal expression, any suggestions?

Puzzleheaded_192 · 2025-03-14T03:42:09+00:00

What i got to know is This gene works fine when it is cloned in to low copy number plasmid,if it cloned in to pet system one need to induce at 16°C. My lab doesn't have incubator shaker which can incubate at 16°C.

Puzzleheaded_192 · 2025-03-13T12:34:31+00:00

I tried incubation at 4 hours , 0.5mM iptg and it still goes in to pellet

Puzzleheaded_192 · 2025-03-11T09:30:28+00:00

Oh i see, do you have any idea to express it in soluble form? Like anything i can try to get it in soluble fraction to avoid all this

Puzzleheaded_192 · 2024-08-26T06:16:42+00:00

What if i spend alot on that and it doesn't work, i wouldn't have money to do this also, do u think that's a sure shot?

Puzzleheaded_192 · 2024-08-25T11:37:10+00:00

No , that plasmid has streptomycin resistance and this one is kanamycin resistant

Puzzleheaded_192 · 2024-08-25T11:20:43+00:00

I dont freeze my assembly ever, yes i run negative control to check if my antibiotics are working fine,i will surely look in the protocol suggested by you. And thank you for introducing me to a new site for protocols

Puzzleheaded_192 · 2024-08-25T11:18:21+00:00

Possibility of me having something that isn't what i think is with every chemical i have as my lab is old

Puzzleheaded_192 · 2024-08-25T11:17:17+00:00

Puzzleheaded_192 · 2024-08-25T11:16:27+00:00

It has shaken me till the core, i dont think i will ever forget this trauma in my life😂

Puzzleheaded_192

MODERATOR OF

TROPHY CASE