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~
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.
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
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🥺♥️
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!
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.
This was explained so perfectly, simply yet in all the necessary detail......THANK YOU so much sir!
I have a Biotechnology aexam coming up soon, this video conteins 15 pages in my book, thanks a lot !
Within 5 minutes clearly understood the concept 😎... Tnkuuuuuuu sirrrrrrr👍
Excellent presentation sir!
From Bangladesh 🥀
Your explanation is very helpful.Thank you 😊
I was always confused about the omega protein, but your video showed me it comes from the chromosomal DNA. Thank you!!!
Excellent execution........
Thanks for such a nice demonstration.......
Thanksss very helpful!👍🏽
From France!
Thank you greetings from Germany
Wonderfully explained. Thank you .
digga.....i mean, bro, this is amazing! very nice animation and explanation! pls more :)
Danke Dir Digga
@@henrikslab de nada ;)
awesome video, i was confused and you clarified everything for me thank you sm!!
Short & precise expl. Thank you so much😊👍
Great!
Thank you sir. Excellent
شكرا الشرح كتير واضح thank you very much for this amazing explanation ❤
it was helpful!! thankyou
Thank you so much for this great explanation
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.
Thankuuuu very much sir......it helped me a lot
Nice explain sir thank you for your help from Al Habib Ahmed from Manipur India
Was very helpful. Thank you
Thank you for your video I understood it more clearly.
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)
Nice 🥳
Thank you so much 🙏
Thank you so much
It was very helpful. Thank you very much!
Thanks !!
This was really helpful. Thank you so much.
Good explanation
It was really helpful. Thank you.
Thanks nd can we write in xeam ??
Great video
danke schoen! (thanks from Taiwan!!!!)
You have to cut subtitle because it is hampering to see the picture nicely.
Really helpful
Thank u very much
Very gud video
Thank you so much.
Thank you sooo much
thank uu so much
Thanks a lote , that was so usefull🙂🙂
thanks
can't thank you enough!
❤
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?
Thanks!
I have exam today thanks for this vedio ❤
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).
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
Nice
Thanks a lot
thanks a lot
Thanks sir
Danke bruder
Love you from kashmir
Tnx
Henrik springer
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! :)
Absolutes alman English
- Renegade musste mega lachen :D schon bisschen true!
thank you so much
Thank u so much
Thanks !