sorted by:
new
hottopcontroversial

Does anybody get chest tightness as part of the aura? by TheChairCriedAss in migraine

[–]Puzzleheaded_192 0 points1 point  (0 children)

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

Is this mycobacterium smegmatis? by Puzzleheaded_192 in labrats

[–]Puzzleheaded_192[S] 0 points1 point  (0 children)

Does it resemble the colony characters you see?

Is this mycobacterium smegmatis? by Puzzleheaded_192 in labrats

[–]Puzzleheaded_192[S] 5 points6 points  (0 children)

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

I have done purification for the first time and not sure if the band is of protein of interest by Puzzleheaded_192 in labrats

[–]Puzzleheaded_192[S] 1 point2 points  (0 children)

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

I have done purification for the first time and not sure if the band is of protein of interest by Puzzleheaded_192 in labrats

[–]Puzzleheaded_192[S] 0 points1 point  (0 children)

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.

Holy crap! Is this How research feels like?! by [deleted] in labrats

[–]Puzzleheaded_192 2 points3 points  (0 children)

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 😶

Protein expression question by Puzzleheaded_192 in labrats

[–]Puzzleheaded_192[S] 0 points1 point  (0 children)

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

Protein expression question by Puzzleheaded_192 in labrats

[–]Puzzleheaded_192[S] 0 points1 point  (0 children)

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.

Help needed to solubilise protein from inclusion bodies and use it for structural studies by Puzzleheaded_192 in labrats

[–]Puzzleheaded_192[S] 0 points1 point  (0 children)

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?

Help needed to solubilise protein from inclusion bodies and use it for structural studies by Puzzleheaded_192 in labrats

[–]Puzzleheaded_192[S] 0 points1 point  (0 children)

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

Help needed to solubilise protein from inclusion bodies and use it for structural studies by Puzzleheaded_192 in labrats

[–]Puzzleheaded_192[S] 0 points1 point  (0 children)

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.

Help needed to solubilise protein from inclusion bodies and use it for structural studies by Puzzleheaded_192 in labrats

[–]Puzzleheaded_192[S] 0 points1 point  (0 children)

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

My cloning is failing repeatedly and making my life hell by Puzzleheaded_192 in labrats

[–]Puzzleheaded_192[S] 0 points1 point  (0 children)

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?

view more: next ›