Hey scientists, I hope you can learn something out of this weeks topic "Blue-White Screening". Thanks for watching! Cheers, Henrik Instagram: / king_henrik_the_1st
Thank you for helping me out by clarifying flawlessly my doubt. This concept was really bugging me and no one was there to help me out . Really grateful to you.
Thanks a lot!! This screening method had confused me for a couple of months and your explanation has helped me understanding it in just 5 minutes!! thanks again~
Fantastic video, personally I was confused about the difference between the "recombinant" colonies and the "transformed" colonies... now I undestrand that all the recombinant are transformed, but not all the transformed are recombinant... thanks you so much!
Shaina C exactly! So you just pick the white ones and transfer them to a different plate. Generally to confirm again if they really have the inserted gene, you do a PCR with some white colonies to genotype (use primers for the insert)
How can we assume that all the recombinant DNA got transformed into bacteria? What about those bacterial colonies who didn't uptake the Plasmid vector, they'll also give white colour. How to differentiate then?
why is it important to know if it's a bacteria with or without a plasmid? Is the plasmid always brought into the bacteria by humans or is it just naturally there?
ThousandFo0tKrutch Good question! By nature bacteria contain small circular plasmids beside their main genome. However, in this case we [humans] brought them in the bacteria (this can be done via heat shock or electroporation where the membrane becomes permeable and takes up plasmids we want them to take up. So that is why we want to know that we just have successfully transformed bacteria: the ones where the heat shock failed are without our dna of interest. Hope that is clearer now.. if not please reply!
@@henrikslab wow thank you very much for your answer. but there is antibiotic in the agar to make sure only bacteria with a plasmid with antibiotic resistence survive. but doesn't a plasmid with this resistence not also always contain the gene of interest? like how is it possible that there are bacteria with a plasmid and a restistence but without the gene of interest? because only these types of bacteria would then show up as blue colonies right? without a plasmid, they would not survive and with the gene of interest the colonies would be all white.
ThousandFo0tKrutch the antibiotic on the plasmid is used as a control whether the plasmid has been taken up or not. Colonies without plasmid might die on antibiotic media as you say. For the living colonies: The blue or white indicates then whether the gene of interest is correctly inserted in the plasmid that was taken up. To your question how it is possible that bacteria have a plasmid but no gene: when you clone the plasmid you might not correctly insert the gene of interest (then you have an ´emptyˋ plasmid with the antibiotic resistance but no gene
Наука
Thank you for helping me out by clarifying flawlessly my doubt. This concept was really bugging me and no one was there to help me out . Really grateful to you.
Out of the four videos i watched to fully understand this concept, yours has been the most understandable for me. Thank you!
krazed98 so nice to hear that! Thanks!
Thanks a lot!! This screening method had confused me for a couple of months and your explanation has helped me understanding it in just 5 minutes!! thanks again~
Me too🥺♥️
This was explained so perfectly, simply yet in all the necessary detail......THANK YOU so much sir!
Excellent execution........
Thanks for such a nice demonstration.......
Wonderfully explained. Thank you .
Your explanation is very helpful.Thank you 😊
awesome video, i was confused and you clarified everything for me thank you sm!!
Short & precise expl. Thank you so much😊👍
This was really helpful. Thank you so much.
Within 5 minutes clearly understood the concept 😎... Tnkuuuuuuu sirrrrrrr👍
It was very helpful. Thank you very much!
Was very helpful. Thank you
شكرا الشرح كتير واضح thank you very much for this amazing explanation ❤
Thank you so much for this great explanation
Thank you for your video I understood it more clearly.
I have a Biotechnology aexam coming up soon, this video conteins 15 pages in my book, thanks a lot !
Thanksss very helpful!👍🏽
From France!
Excellent presentation sir!
From Bangladesh 🥀
I was always confused about the omega protein, but your video showed me it comes from the chromosomal DNA. Thank you!!!
Thank you so much.
Great!
Thank you greetings from Germany
Thank you so much 🙏
Thank you so much
digga.....i mean, bro, this is amazing! very nice animation and explanation! pls more :)
Danke Dir Digga
@@henrikslab de nada ;)
thank you so much
can't thank you enough!
Thankuuuu very much sir......it helped me a lot
Great video
Thank u so much
Thank u very much
Thanks !!
Nice explain sir thank you for your help from Al Habib Ahmed from Manipur India
Thank you sooo much
Thanks!
Thanks nd can we write in xeam ??
thank uu so much
Thanks a lote , that was so usefull🙂🙂
danke schoen! (thanks from Taiwan!!!!)
Thanks a lot
Fantastic video, personally I was confused about the difference between the "recombinant" colonies and the "transformed" colonies... now I undestrand that all the recombinant are transformed, but not all the transformed are recombinant... thanks you so much!
Then how to differentiate between Recombinant and Non transformed colonies? Since both will give white colour.
Nice 🥳
thanks
Good explanation
Really helpful
So the white colonies on the ampicilian plates indicate that cloning was successful correct?
Shaina C exactly! So you just pick the white ones and transfer them to a different plate. Generally to confirm again if they really have the inserted gene, you do a PCR with some white colonies to genotype (use primers for the insert)
Thanks sir
How can we assume that all the recombinant DNA got transformed into bacteria? What about those bacterial colonies who didn't uptake the Plasmid vector, they'll also give white colour. How to differentiate then?
Very gud video
Tnx
Nice
You have to cut subtitle because it is hampering to see the picture nicely.
❤
So what I am confused about is how are there both blue and white colonies, do the blue ones just failed to pick up a DNA of interest?
Yes, either that or they did not pick up the whole plasmid (/vector).
Danke bruder
I have exam today thanks for this vedio ❤
Wooow i got it well better then my prof
why is it important to know if it's a bacteria with or without a plasmid? Is the plasmid always brought into the bacteria by humans or is it just naturally there?
ThousandFo0tKrutch Good question! By nature bacteria contain small circular plasmids beside their main genome. However, in this case we [humans] brought them in the bacteria (this can be done via heat shock or electroporation where the membrane becomes permeable and takes up plasmids we want them to take up. So that is why we want to know that we just have successfully transformed bacteria: the ones where the heat shock failed are without our dna of interest.
Hope that is clearer now.. if not please reply!
@@henrikslab wow thank you very much for your answer. but there is antibiotic in the agar to make sure only bacteria with a plasmid with antibiotic resistence survive. but doesn't a plasmid with this resistence not also always contain the gene of interest? like how is it possible that there are bacteria with a plasmid and a restistence but without the gene of interest? because only these types of bacteria would then show up as blue colonies right? without a plasmid, they would not survive and with the gene of interest the colonies would be all white.
@@henrikslab also are you german?
ThousandFo0tKrutch the antibiotic on the plasmid is used as a control whether the plasmid has been taken up or not. Colonies without plasmid might die on antibiotic media as you say. For the living colonies: The blue or white indicates then whether the gene of interest is correctly inserted in the plasmid that was taken up.
To your question how it is possible that bacteria have a plasmid but no gene: when you clone the plasmid you might not correctly insert the gene of interest (then you have an ´emptyˋ plasmid with the antibiotic resistance but no gene
ThousandFo0tKrutch I refer to the top comment - I am
Love you from kashmir
Henrik springer
Absolutes alman English
- Renegade musste mega lachen :D schon bisschen true!
Red doch gleich Deutsch, deinen Akzent kann man sich nicht geben! 😄 Is nicht böse gemeint.
Sry, i don´t really get what you mean? Could you write in english please? Kappa
@@henrikslab ehre
Gewöhn dich daran. In der Naturwissenschaft, vor allem in der Biotechnologie, ist Englisch unausweichlich. Ich finde, er hat es sehr souverän gemacht.
@@henrikslab hahahahah
Aber ist echt ein gutes Video geworden! :)
It was really helpful. Thank you.
Thanks !