Insertional inactivation (two antibiotic selectable markers) | Biotechnology | Khan Academy
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- เผยแพร่เมื่อ 29 ส.ค. 2024
- Let's explore what insertional inactivation is, and how it helps in selecting cells that have taken up the foreign DNA from the ones that have just taken up empty plasmids.
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Created by Khadijatul Qubra
8:10
THIS is the exact reason i had to watch all these kinds of videos online after reading this topic from the NCERT! I didn’t understand at all why we are making the transformants if we are going to kill them anyway in Tetracyclin. Thank you sooo much for clarifying this now I know why
lol same
we are killing them for identification purpose ,but can get it back from other replica plates made
1)differentiated between transformant(who took plasmid may be recombinant or non recombinant) and transformant(no plasmid) using ampicillin culture
2)differentiated between recombinant and non recom. (Plasmid which have our gene of interest and that which don't).We put our gene of interest at the place of tetracycline resistance gene and due to this cell became sensitive to tetr and died .
We got that died tetr. Colony back from the replica plates(that colony which are at the same place )
@mitaigharsabahath2918 see, at the end we just got to know about 3 and 4 aur non recombinant so NOW we can remove 3 and 4 from the master plate and hence the master plate now will only have 1 and 2 colonies as 5 was already killed before and we removed 3 and 4. hope it helped😅
Well said@@saraasif7988
Perfectly explained for my revision
that example was on 6:34 was incredible made so much sense
Finally got it after watching it for two times! 😭
Awesome explanation without any lost of time.
I think this is the only video on TH-cam which explains this topic much clearly❤❤
Awesome explanation cleared each and every ncert line
Thank you so much Khan Academy for such a nice explanation 💝
Still I haven't got how in real lab setting they will differentiate between first two types , externally all the colonies will appear same so can not be labelled with numbers?
Hey I got that , I didn't know how they make these photographic plates but may be they do so by transferring some bacteria from each colonies on exact same places as previous petridish or by numbering as you do . Then it's simple to deduct which are which bacteria and thus we got it. Thanks.
Those are bacterias , not an animal with external morphology and a distinguishable feature and they are so small
You can't find exact location again.
I THINK SO 🤔
there's a procedure called replica plating. that's used to stamp all the colonies present on the master plate onto a new plate on the same location.
No one explained like you mam 🙏🙏🙏
One of the best explanation on youtube 😂
Amazing explanation. Thank you
Thank you
Awesome explanation ma'am
So crystal clear. Thanks a lot. Hare Krishna Dandwat Pranaam
Huge thank you i now cleared my doubts
THANK YOU. You are excellent🥰🥰🥰
Mam Please teach 'molecular basis of inheritance' next
Check the website, udhar hai
Thank you so much ma'am
I was confused that if recombinant dies in tetracycline then what is the use of it... Thanks mam ❤❤❤
I had the same question. But now my doubts are solved. NCERT does a poor job at conveying the complete information 😵💫. But hey, happy learning...
Nice explanation 👍
Now I also came to know that some student and teachers done this topic through ratification from NCERT (mine experience)
Bcz when I ask them, if recombinant die , what the use and they🤐
Same bro
Swear on Allah , anytime i need help i come to khanacademy .
You Just explained it the way it should be explained
Thank you so much! Awesome video!
Thank u for clarifying my big doubt 💝
I get that this explanation seems to make sense, but that would only be the case if the colonies form at the exact same positions in the petri dish, no?
Can we really just assume that the colonies 1 and 2 on the first petri dish are also formed at the positions 1 and 2 of the second one?
I don't think so.
Perhaps it might have been better to add a gene that codes for a protein which maybe glows under certain conditions in place of the tetracycline resistance gene (lac z gene, for example). That would cause colonies 3 and 4 to glow, while 1 and 2 don't, due to insertional inactivation.
What do you think?
The duplicate plates are like your clones, you do experiment on them to find out the correct way. Then from the result you operate on yourself. But my point is how do you make those exact clones in the first place. That is my real doubt.
It was best. Thank youu ❤❤
Please upload chemistry videos too..
Apriciate your efforts 👌
Good man good man
Very nice
It would be better if you labelled recombinants , transformants etc
Thank you so much
Woow nice explain and clear voice😎
Best explanation
Thank you!
Superb
Thanks
Still I am confuse because why we kill recombinant DNA carring bacteria(1,2) although we need them. Please anyone clear this doubt
Petridish have four quadrants! Which make them know about it !
Is it possible a plasmid has only amplicillin antibiotic resistance gene? If so what would happen if plasmid has only amplicillin antibiotic resistance gene? This will survive in amplicillin but will die in Tet. like the recombinant gene do? How can we seperate these two.
🔥🔥🔥
What is the name of mam anyone knows???? Tell me
Why jerk