What is Insertional inactivation? How it helps in the selection of recombinant colonies?

[@vanshikahere1304]

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[@biologyexams4u]

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[@miss_potato1312]

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[@rakhi3836]

The best explanation for the topic👌👌

[@addyfluence183]

But how we will separate the transformed with recombinant vector containing bacteria from transformed with non-recombinant vector containing one... from the tetracycline plate.. As both have survived in it?

[@advythgoud4664]

For that we need to continue further with replica plating. Once when both the recombinant and non-recombinant transformed cells have formed colonies in the Amplicillin plate, using replica plating technique we place the same colonies in the Tetracyclin plate. Then observe which colonies have not grown. Based on the absence of the colony position, take the previous Ampicillin plate and select the colonies. Those are the transformed cells with recombined Gene.

[@aspirant2385]

Remember Transformed with non recombinant would grow in both resistance gene ( ampicillin, tetracycline) but the transformant with recombinant would lose 1 of the property of resistance gene where it had been linked to the plasmid . For example if you link a gene of interest(alien dna) at BamH1 site of tetracycline resistance gene the transformant with recombinant would only grow in ampicillin medium and lose its ability to grow on tetracycline medium whereas transformant with non recombinant would grow on both ampicillin and tetracycline medium

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[@biologyexams4u]

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[@vivekguatam1551]

My doubt is that when we transfer recombinants along with non-recombinant bacterial cells to tetracycline containing medium. The recombinants cells should die as the tetR gene in these cells have been inactivated. While the non-recombinants which contain both the ampR and tetR genes should survive in both ampicillin & tetracycline mediums. So how do we separate these recombinants & non-recombinants cells from tetracycline containing plates as recombinants here should have died while non-recombinants have survived?

[@aspirant2385]

Thank you sir all the doubts regarding insertional inactivation had been cleared with your help

[@Raptor9323]

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[@jagapradhan552]

Excellent explaining👌👌👌

[@venkataramanen]

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[@jaigupta6034]

really very clear explanation

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Brief explanation Sir . Thanks for clearing my doubt ! Keep making videos ☺

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[@himanshusuryawanshi1031]

Awesome explanation

[@gulamjilani9090]

It was like reading a script buddy .....plz explain what are transformants and recombinats

[@pahulsingh1672]

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[@Sneha-gb9ep]

very nicely explained....:)

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[@yashwantkush]

What are these red green circle represt

[@anushanathan1419]

very short and understandable tku

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awesome explaination

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really helpful!!

[@ManjunathManjunath-xm3pl]

Thank you very much helped a lot

[@zuhrab00]

Well explained😍😍

[@biologyexams4u]

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[@pravallikakarri3473]

Oh..so later the inactivated recombinants are used in other processes ah? Am i right sir? Or plz tell the correct ans

[@biologyexams4u]

Yes... Inactivated recombinants means that contain our vector with our gene of interest.. or insertional inactivation has happened due to insertion of our desired gene... Yes then colony is taken for further process as it contains our gene of interest. Hope you are clear Thank you

[@pravallikakarri3473]

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[@faizm7807]

Thanks 😙

[@supriyamajumdar7346]

Damn gud sir...Thanks a lot... Really appreciating one

[@ansarinaziya9715]

How master plate is formed??????

[@akiztoi]

Hi, What if we have some transformed bacteria with recombinant vector but without our gene of interest? For example when we use BamHI to cut a big molecule of DNA which conatains a little fragment (gene) of interest for us? How could we differentiate recombinant bacteria WITH and WITHOUT our genen of itnerest? Thanks and great video!

[@pckunhammedlic551]

Usefull😊👍

[@booya4ah]

THANK YOU SO MUCH

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[@shivani2065]

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[@AnjaliSingh-wi9ox]

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[@shahzadpoetryandbiology7512]

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[@ibrarshahswaty745]

Fast speed😣

[@biologyexams4u]

Hi Ibrar Sorry for the inconvenience. Hope the video helped your understanding. Will slow down in the upcoming videos. thank you for the suggestion and support:) @biologyexams4u

[@rifathmagsum5115]

U can slow down perhaps