Designing gRNA Oligos to Clone into Cas9 Expression Plasmids for KO Experiments
Вставка
- Опубліковано 28 сер 2019
- Description of the steps required to design effective gRNA sequences and then clone those sequences into a Cas9 expression plasmid for KO experiments.
- Навчання та стиль
I can’t believe someone can explain this so simply while being detailed! Thanks Elmer! 👍
I read 200 papers to understand CRISPR knockout and it was almost impossible. This is the best explanation ever. Simple and efficient. Thanks!
I am usually skeptical of long videos, but this one explained everything I needed to know well and did not waste a single second. Thanks!
such a great videos i read so many papers but nobody explained this clearly
Thank you soooo much! This video is extremely helpful. Should mention that you have a nice voice and pronunciation, it's so important for a non-native speaker:)
this video much more informative and clear than classical procedures, especially the chopchop part is very helpful. . Thanks a lot for sharing.
I paid fees to people that confused me. In less than 30mins you explained everything. Thanks so much.
This was really helpful, thank you for making it!
Probably the best video..... your efforts are really appreciated:-)
Thank you so much for the simplicity and clarity of the instructions
Thanks so much! This was incredibly helpful and I appreciate all of the useful tidbits!
The greatest explenation of CRISPR in UA-cam. Congratulations
This video was amazing, Jacob! thank you so much for sharing it!
Thank you so much. The way you explain by breaking everything down makes it so easy to understand. I hope you make more videos like this. Also, your way of articulation and slides are amazing and never bores me out.
This is the best video I have found on this topic. Thank you.
thank you for making this video in such clear and practical way!
It's just the perfect explanation. Thank you!
Very informative especially for the beginners. Thank you for making this video!
Very informative lecture with details, thanks a lot!
Dude, this is thy best CRISPR video on the net! tnx
cant thank you enough for making this video!
Great job, Jacob!
Thanks for this very explicit video
Best tutorial on this topic, I've ever watched. You have another subscriber here. Many thanks!
So beginner friendly and helpful. Thank you so much! ♥️
Thank you so much! very useful and clear explanation!
OMG!!!! FINALLY, I GOT IT! This video is the best ever Jacob:) Thank you
Grateful for this video. It has been helpful. You are a good teacher.
Extremely helpful! Thanks a lot
Congratulations Jacob Elmer, I really like it. Super didactic. Surely one of the best video.
More than a Perfect Video!!
thank u for simple but nice explanation, it helps me a lot
Thank you so much, you really explained so clear🙏
Amazing video easy to understand and you have a really nice way of explaining ! Keep up the good work
Thanks a lot Jacob... .. . Even few of my Postdoc Colleagues were not able to make me understand it like the way you did... .. . Really appreciate it a lot
I love your explanation..
Thanks thanks thanks for the help! You're a legend!
i have never done CRISPER, after watching this great master lecture i am confident that I can do it very well in lab. very detailed practical steps.
Sir, very well informative lecture for me I loved the way you explained.
Thanks for the wonderful informative video :) .
Great video man, my professor just isn't all there and no one in our course understood how to design a gRNA :/ With your Video I just get it now. Thanks for the clarity and especially the explanation how the algorithm works and how you could do it yourself. Such a great help thx again
Perfect explanation thank you!
Oh my God, you save my life. Thanks a lot!
This is great! Thanks!
Comprehensive details, the Best method of explanation. I am gonna subscribe and like your channel to support your generous efforts. Thanks btw for the time and effort you put into this video.
very good video. Easy to understand such a difficult approach.
Thanks jacob. I loved how finely you explained everything. I was wondering if you make a video on crisprs mediated repair outcomes.
Really nice tuto thanks !
This video is amazing
Thanks a lot for your explanation it helped a lot
Amazing explanation
Utterly resourceful, definitely deserve more views and subscriptions!!
Btw, chopchop is constantly in use, priority 6-9 is common..
Very informative video. Clear all doubts about gRNA design.
awesome video!! thanks!!
I have been looking for a good source for gRNA design for several weeks. Only with this video, I could find the answer to almost all of my questions. Thanks a lot, Jacob. Great job. Would you please add a quick video on how to design gRNA for CRISPR/dCas9 system for gene expression regulation? Thanks
thanks a lot, it is extremely informative !
Thank you. Please make more videos on techniques used in the field of Biotechnology. The way you explain procedures and processes is very easy to understand and follow. You, sir, are amazing.
This is outstanding
thank you very much, it's very helpful!
Very useful and instructive
Very useful contents.thanks dear
This was very helpful
So nice presentation... Very informative & easily digestible material.. Much useful than other Videos.. Thanks a lot
thanks for this tutorial.
SIMPLY PERFECT
Awesome 🎉
Thank you!
Awesome Video thanks alooooooooot
Thankyou so much! Please keep making engineering videos
great video indeed
Very nice dear sir
Really helpful. More video expected on CRISPR
Te amo, gracias me salvaste
AMAZING THANK U SO MUCH
Bravo!
sir u are amazing
I really loved the way u explained
u made my job much easier
plz make an video how do I clone a dna segment in to a vector in detail
Genius!
My first word after watching this video was WoW. Not usual for me.
Dear, Greetings, You are awesome
the best video! thank you, please can you continue this video for the next step of CRISPR/cas9?
good work
much love
Superb
Lovely
Great video, thanks for taking the time to explain how it works, this is great help. I have one question, how should I proceed if I want to excise a potion of an intron and remove it completely? Thanks in advance.
Thank you,, Please make a video on Analysis of ON and OFF target from NGS seq.. Cheers
Such a clear and informative video! I had a quick question. I am looking to knock out a gene cluster in bacteria. Would this procedure work in bacteria as well?
Thank you very much 🙂
That was a great video. Thank you very much. How about if we want to Knock in a gene?!
Informative! Can you also make a video about knock-in design ? Thanks.
Hi Jacob, I am glad I found your video! It is really helpful. However, I still have one question: if you have these sticky ends between gRNA and scRNA, how do they assemble correctly?
I found your video more than useful! Thanks a lot...and I have a question! there is a way to see in what exon is located targets sequences ? I mean, not manually checking every exon in the gene... If you work with a big gene, with lot of exons, it can be pretty annoying look for your target sequence just checking sequence along.
Yes use snapgene software
thanks a lot,... so well explained , could you please explain more about NGG sequence ? thanks
basically its the PAM sequence and I think its used as a way to identify the difference between the gRNA and the sequence
A wonderful and informative video. I had one quick question, I'm looking into knockdown of a gene (INPP5D) in human genome but am wondering the main differences between the two options for homo sapiens provided by chopchop. If you could clarify the significance that would be very helpful. Thank you!
Are you referring to the hg19 and hg38 versions of the human genome?
Hi Jacob! very helpful video. Thanks. Anyone has any idea about how to make a plasmid that induces double KO for 2 genes? Or better to say, how to make a double KO cell line.
Thank you, the video is a great help; but what about the scaffold RNA? is it included in the plasmid in addition to Cas9 encoding gene?
In this case, we do not design the plasmid, we purchase it from some company!
thank you so much sir. It really helps me a lot. Is it possible for you to share the power point? through Gdrive may be?
Hey this video is super helpful and well made! Do you have any supporting literature you could provide?
Very informative...have a little query...what that + written means ..is this target sequence found in coding strand or whether it will be targeting the coding strand..?
Dear Jacob, thank you for the nice lecture! I wonder, is there a company that if you seen the gRNA they will send you already the cloned plasmid?
i really liked your video..can you help me in designing the sgRNA for MLo-1 gene of Triticum aestivum??