Synthesizing cDNA with Reverse Transcriptase
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@TheDuvee6
6 жыл бұрын
he's very professional
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@eintroll8792
2 жыл бұрын
Excellent indeed, excellent in spreading wrong information.
Thank you for existing Sir, this helped a lot
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@AKLECTURES
Жыл бұрын
Damn it was really 8 years ago?
@emirkaplan4621
Жыл бұрын
@@AKLECTURES time is passing mister, you did absolutely well. We are-mbg students- still watching your videos. Thank you
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Fantastic Overview of this process.
@AKLECTURES
9 жыл бұрын
Damian Clark thank Damian! :)
@116chandra
7 жыл бұрын
AK LECTURES Tools of biotechnology
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@QuentinWolffMusic
7 жыл бұрын
But I have a question: The gene is different at the end, there is a new codon (CCC/GGG) which can produce another protein, isn't it a problem ?
@mrinaliniroy8496
7 жыл бұрын
+QuentinWolffMusic no it won't be a problem because the translation of the mRNA formed from this cDNA would have a start and stop codon flanking the gene of interest, so any no. of dNTPs before the start codon won't alter the final protein. I hope ths,helped
thank you so much. with your clear explanation i start to love molecular biology. ;)
6:08 A poly-C tail should be a poly-T tail.
Great job explaining this! couldn't be any clearer!
@juliabl6471
7 жыл бұрын
No, DNA is not single strand. What we do first is to create the ssDNA complementary to the ssRNA. We now have to destroy the ssRNA to build the complementary in DNA. That is why we use high pH to eliminate RNA and only obtain ssDNA. Then with DNA polymerase the second strand of DNA is created. Now we have dsDNA. Nothing of this means that DNA is simgle stranded, only when we need it to be in the process using heat.
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Thank you soo much! you are genius :) So helpful
So helpful video, thank you so much
Very easy to understand!
Thanks sir.......you are genius
brilliant ! thanks a ton :)
@AKLECTURES
9 жыл бұрын
Gayathri Jaikumar you're welcome!
Great video, thank you :)
Very useful... thank youuu !
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Excellent video!
@AKLECTURES
9 жыл бұрын
Nahu Dimitri Thanks! :)
very great thank you.
Thank you
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thank you
well explained
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Silly question, do humans have reverse transcriptase or is that only found in certain viruses
in step number 6 the 5prime end consists a hydroxyl group,do a 5prime end consist hydroxyl group or phosphate group?
You are a medical hero! Thanks a lot!
sir u r the best....ur teaching method is amazing...so easy to understand
There is an error at 2:25 ( you said "exons that must be spliced out", when it is Introns )
Very good 👍
exons that must be spliced out? I thought introns, as you said in the beginning? :/
@gerardosaa9396
6 жыл бұрын
he just said it the other way. but he puts the concepts so clear that no one notices it
Your video has a mistake. You said that for the eukaryotic mRNA, the exons must be spliced out but it's the introns that are spliced out during the post-transcriptional modification to produce the processed mRNA. time 2:30
@urselkhan5868
5 жыл бұрын
That's not a hard-fast rule. mRNA can undergo "Alternative Splicing" where exons are either spliced out or left in the mRNA. It is based on the splice site that the spliceosome decides to function at.
great vid!
@AKLECTURES
9 жыл бұрын
Artur livshits thanks! :)
How can we know that the poly T tail won't bind to the poly A tail of another mRNA molecule rather than the mRNA molecule of interest since all the mRNA molecules have a poly A tail
@sebastiaanbol5778
6 жыл бұрын
You don't. Poly T-tails will be used for non-biased cDNA library generation. If you want to make gene specific cDNA only, you will need a 3' reverse primer that is sequence specific.
A question, E. Coli can produce RNAm eukariotic? What happen with the rbs of the eukariotics cells? In your video we watch that.
thank u sir
Awesome
Why do we need to synthesise poly c tail dna primer?? We can use poly a tail dna primer it is complementary to the cDNA.. And we don't need to add terminal transferase
how are sticky ends attached to the two newly generated cDNA?
Maybe already have long time this video but I have a question, and I hope someone can answer me. There's no technology to create the protein using our eucaryotic mRNA and to avoid the other steps to get a protein (eucaryotic)? I mean, if we can extract the mature transcript (modified mRNA -3' poli A tail, 5' cap and spliced exons) from a eucaryotic cell, why we need to form, make or any word you want to use, cDNA and then use procaryotic cell?, unless there's no technology to use directly the mature mRNA. Thanks
@cubsfan708
6 жыл бұрын
because retroviruses can insert specific DNA fragments into the host original DNA, this means everytime the host cell reproduces it will have this new modified DNA which can be translated in the new RNA , if you directly insert mRNA it will only be used once because mrna degrades over time
Excelent video, but shouldn't the 5' ends of the molecules have a phosphate group instead of a hydroxyl group?
@mmaking8664
7 жыл бұрын
i was wondering the same thing
DNA polymerase isn't needed to finish synthesizing the ds-cDNA molecule. RTase has DNA polymerase activity.
Thank you so much. And RACE PCR please!!! :)
Upload a video explaining adapters and linkers please
Only partially digest RNA with RNAse H, not the entire RNA
Biology jargon! I love it...lol
Page 43 4.7.'76 3.7.'76 From : Adams, J. To : Adams, A.
@nurlatifahmohdnor8939
2 жыл бұрын
9.4.'22 = 7.9.'43 We bought Ak_k in P. C. for our 1st son.
This is graet😎
since the revierse transcirbatse need a primier to start doing its jop whats if we inserted a normal dna polymerase instead of the RT wouldnt it do excatly the same process of making complementry dna to the mrna ?
@TaiNguyen-tw1gp
5 жыл бұрын
I don't think DNA polymerase will do its job on a single-stranded mRNA, as it cannot use the mRNA strand as some sort of 'template strand' (mRNA strands contain uracil instead of thymine). So that is why reverse transcriptase is used