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Which protein you are expressing and purifying

U have used dnase before sonication.. does the addition of dnase before sonication impacts lysis differently or it does not affect the lysis

On original is omeletu formage

It extends 3' end 😂

😂😂My type of content. I'm home now 😅

Thank you !!!

Can you share some references you use to do in this video? Thank you so much

Thanks for the video. As a first-year PhD student, I needed this video to get my feet on the ground. Thanks a lot

All Biochemistry Master's Students looking to understand the method of protein expression for your project hands up ✋🏽

Hi where is your lab I have a few questions

School crushes be like

Please tell me how you made your elution buffer, everytime I add imidazole the pH rises above whats needed. And adjusting it with acid is not possible because of possible interference.

It is always better to add lysozyme AFTER resuspending the pellet.

🤠

lmao 5' to 3' always!

Great performance, great explanation. Thank you.

Quality content

first of all - thanks, it really helped me to understand several steps of this procedure better! secondly, it's the best unintentional asmr i've heard in my entire life, please, continue

So helpful, thanks a lot!!

love this video thank you!!

10:40 for Day 4

I'm glad that I found your channel. It's really a nice and detailed Protein extraction video on TH-cam that's really helpful for beginners like me 😃

How do you determine what gradient of SDS-PAGE gel to use since there are multiple products on the market that range from 4%-12, or 4%-20% gradient?

Really good and well explained 👍

Hi Great Job Can I have this protocol in written form So i cannot miss any point. It would be highly appreciated

I appreciated the level of detail you described here, thanks. Using your technique, about how long does it take you to grow and purify a 1L culture?

What is the success/trial ratio of this process, if i follow the process exactly, will i be able to express the protein or does it takes few trials

Great video! Great technique. Thank you.

Hi, thank you very much for your excellent job! And I have a question, at 1'43'', what is the work frequency of the machine? Thank you！

whats the name of the spectrophotometer machine you were using?

Hi, this video was great!! Maybe you can do additional videos on SDS-page interpretation (maybe with different proteins and conditions).

LB should be pH'd to 7.

I want to see how the running gel looks like.

thank you the videos ,i am doing sds page but my run wont start even after adding running buffer to the mark .i use the same mini gel tank like yours ,though it leaks ,could it be the issue ?help me from this confusion please

Thank You so much sir

Outstanding sir

Hello! My name is Maria and I am a PhD student working on a project that works to collate various biological techniques for early career researchers in the lab. I really loved your tutorial video for protein purification, and was wondering whether we could get in touch to discuss it further and other similar protocols. Please let me know I'd love to hear back from you!

Thank you very much, this is great for teaching with limited lab.

Aabsolute master! Thanks my dude - this helped a lot! For the viewer's sake you might include on-screen stats for the reagents used. Thanks again! :D

I liked the cotton idea :) on gel staining part of the protocol. Useful indeed

Do you have a protocol please

Please continue this, it's definitely a channel worth subscribing

I do protein expression with 6 liters of culture. After centrifugation of the lysate, the supernatant is passed onto the Ni-NTA agarose column. It takes several days for the supernatant from such a large culture to pass through the column or immediately by syphoning the supernatant out of the column by applying a vacuum to the base of the column.

please write the name of manufacture Ni-nickel resin

Thank you

Is the ice important and why?

Thank you so much for this!

Ahh this was such a joy to watch. I am currently doing my master's thesis on recombinant production of some protein and watching you do this whole process was such a cool experience 😂 I am so motivated now for my GST affinity purification tomorrow 😂💪🏼

May I know what does it mean by to wash with 10 column volume? I encounter this in an article