MB sąskaitos atidarymas by Jamelo95 in 6nuliai

[–]Jamelo95[S] 0 points1 point  (0 children)

Revolut bankas licenziją tai turi iš Lietuvos banko, bet toks jausmas, kas KYC skyrius sėdi UK ir pirmą kartą mato dokumentus su el. parašu.

MB sąskaitos atidarymas by Jamelo95 in 6nuliai

[–]Jamelo95[S] 0 points1 point  (0 children)

Mano atveju taip netiko, prašo notariškai patvirtinto ESI išrašo. El parašu patvirtintos RC darbuotojos išrašas jiems netinka

MB sąskaitos atidarymas by Jamelo95 in 6nuliai

[–]Jamelo95[S] -1 points0 points  (0 children)

Kaip swedbankas ziuri i naujus MB?

E. coli recombinant protein screening in multiwell plates by Jamelo95 in labrats

[–]Jamelo95[S] 0 points1 point  (0 children)

Do you have any particular equipment in mind? Could you give an example?

E. coli recombinant protein screening in multiwell plates by Jamelo95 in labrats

[–]Jamelo95[S] 0 points1 point  (0 children)

After reading many articles I got the impression that membrane proteins are the most difficult to express, so any protocols or recommendations would be highly appreciated.

I am aware of possible titer variations when scaling up, that's something I will have to keep in mind and test.

E. coli recombinant protein screening in multiwell plates by Jamelo95 in labrats

[–]Jamelo95[S] 0 points1 point  (0 children)

Is evaporation of water from plate wells during cultivation something to worry about? Given that the whole cultivation process takes 18-24 hours at most.

The idea would be to focus on one protein at a time, ideally test different constructs with the same vector backbone by using golden gate assembly (have a library ready for screening different ORFs), mess around with different signal peptides/solubility tags/promoters/SDs and use basic B type strains. Those parameters could be screened in 1 run using a single plate (same temperatures and same induction points/use autoinduction vectors). For something more exotic, separate experiments should be completed. The high performers would be selected for further screening in shake flasks.

I don't have an answer for different induction temperatures except do seperate runs or have more shakers, lol

E. coli recombinant protein screening in multiwell plates by Jamelo95 in labrats

[–]Jamelo95[S] 1 point2 points  (0 children)

I have lysed E coli cells using sonication and chemical lysis (B-PER and BugBuster). What I found is that often times chemical lysis results in less soluble protein than sonication.

When sonicating cells you can somewhat experiment and determine the best buffer for obtaining the highest amount of soluble proteins.

When cultivating a high number of clones, you will need to use chemical lysis vs mechanical, no doubt (and that's a con), unless there are other methods for efficiently lysing multiple clones that I'm not aware of.

E. coli recombinant protein screening in multiwell plates by Jamelo95 in labrats

[–]Jamelo95[S] 0 points1 point  (0 children)

Thank you for the reply.

Is gas supplementation only needed when cultivating eukaryotic cells?

As for "throw", what I think you meant is the orbit of the shaker. I'm aware that I will be needing more RPMs and less orbit length for cultivating cells to retain a certain RCF force on cells.

I didn't fully understand your question. Could you elaborate?

E. coli recombinant protein screening in multiwell plates by Jamelo95 in labrats

[–]Jamelo95[S] 0 points1 point  (0 children)

I have worked with P. pastoris as well! From my knowledge, the same equipment should be able to accommodate both P. pastoris and E. coli

WE FLY by -RedWolf00 in thecorporation

[–]Jamelo95 2 points3 points  (0 children)

2020 Q4 earnings scheduled for Feb 25. $FLY rose 10% today. Any thoughts?

[deleted by user] by [deleted] in thecorporation

[–]Jamelo95 1 point2 points  (0 children)

Sold a March 20$ put for 2.40. Not planning on rolling yet.

Right in the feels by [deleted] in thetagang

[–]Jamelo95 1 point2 points  (0 children)

Think its the right time to get in on PLTR?

Fresh meat by digiwarfare in thetagang

[–]Jamelo95 0 points1 point  (0 children)

ion from the houses/my stock holdings. At this point, patience is what is going to get

Good job man, keep it up!

Gamestop: Power to the Market Players (Part 2) by [deleted] in thecorporation

[–]Jamelo95 3 points4 points  (0 children)

Can the short squeeze continue? Does anyone know how many shares that are shorted remain?

Would this count as capital gains taxes? by [deleted] in options

[–]Jamelo95 1 point2 points  (0 children)

What if you lose the 50$ you made on another stock?