blue white screening

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Watching this because a little confusion came up with this particular sub-topic. Thanks, your explanation really helped!

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An excellent illustration of the concept

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[@lucasfernandez4616]

I think something is wrong in 3:57 minute.... if the bacteria IS recombinant (that means that has a LacZ genes) the bacteria is going to produce an enzyme to degradate Lactose (B-Galactoside). This enzyme is going to produce that lactose (which is white) converts into blue... because the Enzyme is going to react with the lactose (white) to separate the sugar of the X-Gal producing a blue color.... so the bacterias that has the recombinant DNA are going to be blue and those who has no recombinant DNA are going to be white.... is that correct?

[@kalpitalakshman4538]

No, if the bacteria is recombinant, it means , the foreign DNA we have inserted has interuppted the coding sequence of Lac-z gene so lac-z gene will not translate to give enzyme (β- galactosidase) without enzyme substrate galx won't be broken down to give blue product

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Plz help me what word do you used at 2:55 while talking about multiple cloning site

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Perturbed

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Very nice sir ...very useful

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[@carlosmaldonado967]

In order to disrupt the lacZ gene via restriction digestion, there would need to be restriction site within the lac Z gene. Do we introduce the restriction site into the lacZ gene to perform the restriction digestion? If so, how does this not perturb the lacZ gene? If it does perturb the lac Z gene, then all colonies would appear white whether it was transformed with GoI or not. Can you explain this? Is there a section of the lacZ gene that is unimportant for transcription and thus the restriction site can be added there?

[@AmnaKhan-se5ek]

The restriction site which is introduced in it is called polylinker. It is so small that it doesn't really affect the lacZ gene..

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Thank you this was clear and to the point

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[@m.sivapriya1648]

It's structure or functional based recombinant bacteria analysis sir

[@animatedbiologywitharpan]

I could not undetstand your question , please elaborate

[@m.sivapriya1648]

In this technique of recombinant analysis of bacteria... Bacterial Functional based analysis or bacterial structure based analysis

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Thank youuu so much.. it is so clear..😍

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Perfect explanation. Thank you Arpan! :D

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Bro u don’t explain about growing those rec bacteria in agar plate in presence of x-gal

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Do non transformed host cells can produce blue colonies on x-gal medium???

[@animatedbiologywitharpan]

Cells which are not transformed with plasmid cant be alive in the antibiotic containing media .....so no chance of getting blue colonies which corresponds to them

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slides are not available...sorry

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Mendelian genetics, NMR, bacterial growth curve, enzyme kinetics, cell signaling.

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Please discuss about buffers

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