James Wright is a good teacher. It was almost I was there with him. I can picture the entire process. He also gave details and principles behind why certain reagents are used. Awesome video. Would like it 100 times if I could.
the screen is already done by inserting the antibiotic resistance gene. A color marker would only be needed if the plasmids were a mix of plasmids with the target sequence and plasmids without the target sequence.
1) The optical density of the E.Coli culture during IPTG induction and IPTG concentration? 2)Too much Ni-beads for that volume of the supernatant, so probably there was nonspecific binding of other protein to the beads leads to the tiny bands in elution lane. 3)The safety issue while the use of glass beaker for sonication. 4) no sample loading for wash step as well as beads!!
If you're working with microorganisms near to a flame. Its dangerous wear gloves. Also the flame sterilize very good the surroundings and usually the bacteria had a antibiotic resistence to keep it clean.
James Wright is a good teacher. It was almost I was there with him. I can picture the entire process. He also gave details and principles behind why certain reagents are used. Awesome video. Would like it 100 times if I could.
Great video! and for everyone alarmed by him not wearing gloves, it's very normal in bacteriology research!
Took me 8 weeks to successfully express a protein, it's pretty simple in theory but doesn't always work smoothly in the lab, great video though!
how difficult are somatropes?
That's the truth!
I appreciate the buffer compositions that you overlayed on the video.
Really thank you for the explanation of such a complicated process in a simpler way
Thank you for that great introduction and clear explanation!
Cómo me gustaría que mi laboratorio estuviera así de equipado u.u
Excelente vídeo, me encantó :3
That is precisely what I was looking for, thanks once more.
Great job, wish you all the best with your research.
i clapped when i see a beautiful nice band u got after purification. nice video
Great video! Thank you for the breakdown
Great Video! James, where ever you are, I hope you're doing well. Cheers bud
Very Useful Video , Thanks for this 👍
Thank you for this video! It's cool how you do this in a lab!
on-point explanation!
You just made my day. Thank you so much.
Thank you very much, this was a very good show and tell for the students.
Its a shame these highly skilled scientists don't get paid what they're worth, even as post-docs.
Great Video Thank you!
Does anyone have any good sources for how this is done at the large industrial scale?
Nice video and very well explication! Congrats!
That plasmid is missing a marker needed for individual colonies separation by color: a screening marker, that is in the MCS region.
the screen is already done by inserting the antibiotic resistance gene. A color marker would only be needed if the plasmids were a mix of plasmids with the target sequence and plasmids without the target sequence.
Nice material!
I like video. Interesting . good describe
Hello,Thank you for the video
Thanks for this lab work divulgation
thanks for sharing!
thank you for explaining
Thank you so much
Hi, can we store the supernatant obtained after sonication and centrifugation that has expressed proteins?
Do you also use any eukaryotic system for protein expression followed by purification than bacterial system?
How do you talk so fast :D I was watching this for exam prep, and I had to slow it down :D Thanks for this though :)
THANKS ALOT ❤
thank you so much!
Are these E. coli producing proteins by cytosolic production, Inclusion body formation, secretory production, or excretory production?
Could you please guide cloning techniques step by step
Thank you; it was brilliant
1) The optical density of the E.Coli culture during IPTG induction and IPTG concentration? 2)Too much Ni-beads for that volume of the supernatant, so probably there was nonspecific binding of other protein to the beads leads to the tiny bands in elution lane. 3)The safety issue while the use of glass beaker for sonication. 4) no sample loading for wash step as well as beads!!
Thank you
You’re the best
if the protein isn't expressed, what does that indicate? what mistakes could I have made?
Inducer is not able to remove repressor or other reasons may be that your bacterial does not allow the expression of protein
No gloves 😱
Yea!! Freaks me out! Good video except for that though.
Shilpa Waduwawara 😀
If you're working with microorganisms near to a flame. Its dangerous wear gloves. Also the flame sterilize very good the surroundings and usually the bacteria had a antibiotic resistence to keep it clean.
is it a cell biology lab or biochemistry?
It can be any sort of biological or chemistry lab, I did this exact same procedure in a Biotechnology lab which was part of a school of chemistry.
Why aren’t you weren’t gloves?
gloves?
Wubbywub the First thing that I noticed
Not needed
where's your glove tho.........
not feeling good without the gloves
No gloves?
So uh, what's he cooking?
Gloves dude...gloves!
It’s not safe to do experiments without gloves and goggles