U have used dnase before sonication.. does the addition of dnase before sonication impacts lysis differently or it does not affect the lysis
@WildorobloxianoАй бұрын
On original is omeletu formage
@xkais487Ай бұрын
It extends 3' end 😂
@g4jmx3z2 ай бұрын
😂😂My type of content. I'm home now 😅
@jenifermunozgomez21023 ай бұрын
Thank you !!!
@atpsynthase17984 ай бұрын
Can you share some references you use to do in this video? Thank you so much
@leahmwendwa5639 ай бұрын
Thanks for the video. As a first-year PhD student, I needed this video to get my feet on the ground. Thanks a lot
@yolisamagibile9 ай бұрын
All Biochemistry Master's Students looking to understand the method of protein expression for your project hands up ✋🏽
@jonathanndunguru11956 ай бұрын
🙌
@Jenny-ym5rg29 күн бұрын
holler 🖐
@harshitasharma795410 ай бұрын
Hi where is your lab I have a few questions
@amahrrinsampson3030 Жыл бұрын
School crushes be like
@sjoerdfennema984 Жыл бұрын
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.
@dr.agupta Жыл бұрын
It is always better to add lysozyme AFTER resuspending the pellet.
@bic1349 Жыл бұрын
🤠
@christophercollins2134 Жыл бұрын
lmao 5' to 3' always!
@TheSergeyVlasenko Жыл бұрын
Great performance, great explanation. Thank you.
@elijahfletcher5944 Жыл бұрын
Quality content
@katerynakozyrieva5631 Жыл бұрын
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
@panoskre Жыл бұрын
So helpful, thanks a lot!!
@blanket6863 Жыл бұрын
love this video thank you!!
@Lussid Жыл бұрын
10:40 for Day 4
@sahilseikh9902 Жыл бұрын
I'm glad that I found your channel. It's really a nice and detailed Protein extraction video on KZbin that's really helpful for beginners like me 😃
@spacescience1002 жыл бұрын
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?
@guleena7852 жыл бұрын
Really good and well explained 👍
@ambreenkanwal89792 жыл бұрын
Hi Great Job Can I have this protocol in written form So i cannot miss any point. It would be highly appreciated
@spectator592 жыл бұрын
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?
@amitmaurya2792 жыл бұрын
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
@lulub50592 жыл бұрын
Great video! Great technique. Thank you.
@loaoo5872 жыл бұрын
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!
@MohammedAli-bj9jk2 жыл бұрын
whats the name of the spectrophotometer machine you were using?
@Chickynugget22 жыл бұрын
Hi, this video was great!! Maybe you can do additional videos on SDS-page interpretation (maybe with different proteins and conditions).
@jinty12322 жыл бұрын
LB should be pH'd to 7.
@shawnbai95432 жыл бұрын
I want to see how the running gel looks like.
@mudondojoyce30902 жыл бұрын
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
@C-Wam2 жыл бұрын
Use fresh running buffer, ensure you have the electrodes the correct way, ensure no leaks, make sure gel is entered the right way, remove the white tape at the bottom of the pre-case gel if you are using those
@hitkarshkushwaha24342 жыл бұрын
Thank You so much sir
@hitkarshkushwaha24342 жыл бұрын
Outstanding sir
@falalalalamyohmy2 жыл бұрын
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!
@hesnayigit88402 жыл бұрын
Thank you very much, this is great for teaching with limited lab.
@kristoffersoelmark6742 жыл бұрын
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
@tinasheprincemaviza752 жыл бұрын
I liked the cotton idea :) on gel staining part of the protocol. Useful indeed
@yordanostselasi45502 жыл бұрын
Do you have a protocol please
@mna1592 жыл бұрын
Please continue this, it's definitely a channel worth subscribing
@adronung18922 жыл бұрын
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.
@C-Wam2 жыл бұрын
Use a 5 mL HiTrap with an FPLC system
@suraalbermani6212 жыл бұрын
please write the name of manufacture Ni-nickel resin
@benysmart16432 жыл бұрын
Thank you
@Stronger.1192 жыл бұрын
Is the ice important and why?
@user-dr9jh6rp6y Жыл бұрын
To avoid protein degradation
@zodeirefo22212 жыл бұрын
Thank you so much for this!
@aleksandar20462 жыл бұрын
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 😂💪🏼
@user-zd7ns9ij5g Жыл бұрын
Your whole MS thesis was on recombinant protein production?
@aleksandar2046 Жыл бұрын
@@user-zd7ns9ij5g and its characterization and potential application in serological test development. But yes, the central part of it was recombinant production of proteins. Sounds pretty underwhelming, right?
@thaborolffy772110 ай бұрын
not really, some proteins are difficult to purify, talking from experience, especially uncharacterized proteins. @@aleksandar2046
@inastasia48712 жыл бұрын
May I know what does it mean by to wash with 10 column volume? I encounter this in an article
@kwanlab40342 жыл бұрын
By "column volume" I'm referring to the volume of resin inside the column, so if there is 1 mL of resin, 10 column volumes is 10 mL.