polymerase chain reaction Animated

[ilhsfm]

this song makes PCR so much more epic haha

[runningbladez]

This makes my life so much easier to understand it :) thank you

[piarocio2013]

@JackBeNimblest We waatch your video all the time in biomedical science class! lol thanks for making it easy to understand. :)

[moos209]

I have no idea what im looking at. Thanks anyway

[cierafe]

simple and straight to the point!

[swifel1k]

Appropriate song, lol!

[MrMike00722]

For some reason, when you add "Let it Rock" to this complex biological technique, it just makes everything easier to understand. Weird.

[GAIArp]

thanks to your help I was able to understand the PCR, thank you!

[IRONMercury]

Whats the difference between TAQ polymerase and DNA Polymerase?

[2madhuV]

@IRONMercury TAQ is a specific DNA polymerase (there are lots of others). It's used more often and more commonly because it's more heat stable, and can resist denaturation when the temperature is increased to induce stand separation.

[SubhanZombie]

Can anybody please tell me whats that 5' and 3' ?

My teacher did that chapter, but I didnt understood anything :P

[SabzoM]

@SubhanZombie 5' means 5 prime end...and 3' means 3 prime end..they are the two ends of the DNA molecule.

and DNA is synthesized by adding nucleotides to the 3' end, so we say that DNA is synthesized in a 5' to 3' direction. since DNA is double stranded and anti parallel, then the opposite strand will be read in a 3' to 5' direction....

hope that helped!

[SubhanZombie]

@SabzoM Thanks a lot buddy :) 

[Yesiquita101]

im loving the song! Great video! Helped a lot! :)

[KostaLaskaratos]

enzymes are fucking crazy, Creation baby. Fucking Creation, this Vid is turning me on.

[stringlessdroid]

@KostaLaskaratos yeah ita fucking denaturing my inhibitions

[kevtyan]

highered(.)mcgraw-hill(.)com/o­lc/dl/120078/micro15(.)swf

Dr. Zeesha eh?

[crazymusicful]

Thank you for making the video :)

[purpleglitterrox]

LOVE saved me from failing

[cubixguy77]

Even taq is shaking to the beat!

[frshVOCALS]

very helpful :)

[Anzu21]

Thanks so much! this video was very helpful to see how exactly it is played out!

[1robyn5]

Not very accurate, is it? I don't think 'cool' and 'heat' cut it..

[The7whoate9]

@1robyn5

What else would you say? He doesn't give the specific temperatures, but this is a good way to visualize it. Also, Enzymes are fucking sick

[zubair908]

@The7whoate9

i just post what i learnt

i dont know the whole topic init

its my required topic

better is i suggest you to ask your professor

[hatedrootofmalice]

@1robyn5 for desnaturalization is at 96 °C, alineation is at 50 -60 °C and for the extension is at 72°C. Aprox.

[sabagul24]

Its jux an awsum way 2 teach... i guess!!! :) gud job..

[sabagul24]

its awesum.



[OldaurGold]

like don't paki make i when video

[ilmaabbas]

this is awesome

[JackBeNimblest]

What happens if you're missing one Amino Acid or have a AA-Pool that is very irregular?...

[zubair908]

@JackBeNimblest  see im also a student..... nt teacher.... so im sorry dude...

[edjorg]

@JackBeNimblest PCR uses nucleotides, not amino acids (aa's are the building blocks of proteins, not the genes that encode them). If one nucleotide is missing then the whole thing stalls on the first round of replication (unless it's an artificial DNA sequence without that base pair- nothing that would code for anything).

[arepeace0687]

@edjorg correct...a.acid is the building blocks of protein...

missing nucleotides in the target gene would not suppress the PCR reaction, unless the missing/mismatches of nucleotides resides in the primer sequences...that could be a problem..

[vallybear7]

@JackBeNimblest What happens is that there is either a Mis-Sense mutation-where one amino acid is substituted for another, or you get a Non-Sense mutation where it can now read a new amino acid calling for the stop, another is the Silent-Mutation which changes the DNA but not the amino acid, hence the name silent mutation, lastly if there is an amino acid removed or added it is called a Frame Shift mutation which causes several amino acids to change.

[vallybear7]

@JackBeNimblest What happens is that there is either a Mis-Sense mutation-where one amino acid is substituted for another, or you get a Non-Sense mutation where it can now read a new amino acid calling for the stop, another is the Silent-Mutation which changes the DNA but not the amino acid, hence the name silent mutation, lastly if there is an amino acid removed or added it is called a Frame Shift mutation which causes several amino acids to change.

[swiftedg]

@JackBeNimblest Amino Acids are not involved in this process, in fact it looks like you may be mixing up replication with translation (in which AAs are associated with tRNA) :) if however you're missing free nucleotides or have an off ratio of nucleotides, you will find that your PCR reaction will have some strands that are incomplete or just not in the quantity that you might want in later cycles. Just remember the central dogma of molecular biology.

[rbrtchng]

@JackBeNimblest You don't need amino acids. This is a gene (DNA), not a protein. If you use an RNA, then you first use reverse transcriptase to get cDNA and use this PCR technique to amplify the cDNA.

[Kiragano]

@JackBeNimblest

this is dna not enzymes...

[smeeth1000]

@Kiragano It's both. Taq polymerase is an enzyme

[lekichnhuong]

@JackBeNimblest I think DNA is sequence before doing PCR (to know what primers to use) => AA pool should not be a problem. However, if a aa is running out, i think polymerase will not going forward since it will see it as a mistake and will not proceed.

[arkhumrox]

@JackBeNimblest I think they put an excess of amino acids just to be safe, but that's just a guess :)

[PND3M1KR3V0LUT10N]

@JackBeNimblest @JackBeNimblest If youre missing an amino acid, translation is affected, not replication. If you meant "missing a (dNTP or nucleotide) then youll have incomplete replication of the target DNA sequences. You will get far fewer copies of the DNA because once the missing dNTP comes up in the replication sequence, replication cant continue. That missing or low-concentration dNTP is the limiting reagent in the reaction. Missing codons = mutations~~>cancer or other disorders

[PND3M1KR3V0LUT10N]

@JackBeNimblest Sickle Cell Anemia, Muscular Dystrophy are examples of non-cancer disorders that arise from mutation in a DNA sequence. Sickle Cell Anemia comes from the replacement of one codon with another, typically by the replacement of one base in the three-base codon sequence (i.e. DNA : ATT --->CTT corresponds to AUU ---> CUU RNA sequence which would replace an isoleucine with a leucine (Amino acid) The way the R groups of AA's interact determine how the protein product folds & functions

[PND3M1KR3V0LUT10N]

@PND3M1KR3V0LUT10N  If the protein doesnt fold right, it doesnt work right, and problems start at the cellular level, which can be compounded by other parts of the cellular machinery not working in response to the first malfunction, which can cause issues at the organismal level.

Cystic Fibrosis is an example of a disorder caused by a mutation in the gene for a type of chloride channel protein in the lungs.

[tarabowers1]

@JackBeNimblest concentration of the "amino acid" pool is actually a pool of dNTP - or nucleotide precursors, if there is an irregularity in these pools you may end up with mutations or mistakes

[silindilemyeni]

My lecturer made us watch this in class...ppl just got up and started dancing.But yeah it totally makes more sense here!!!

[kloepser]

great job!

[screamscream12345]

awesome! simple and effective :)

[isallt]

ye thanks, its quite simple, i found this helpful in telling me where the primers go and now understand how there can be pure target DNA and "dirty" target DNA :)

[alejoperez001]

I love this video, i understood everything and the music is great

Thanks!