The principle of PCR-Polymerase Chain Reaction, a full and easy explanation

This video explains completely and easily PCR, the technique, the principle and the protocol.
If you want to know more about DNA synthesis, press the link below:
• A full explanation abo...

[@RohitPant04]

*Why DNA synthesis occurs in the 5'-3' direction?* Since DNA polymerase requires a free 3' OH group for initiation of synthesis, it can synthesize in only one direction by extending the 3' end of the preexisting nucleotide chain. Hence, DNA polymerase moves along the template strand in a 3'-5' direction, and the daughter strand is formed in a 5'-3' direction.

According the Kary Mullis (inventor of the PCR technique), viral and bacterial infections aren't valid applications for PRC. Why then is this application listed here?

[@a.larson2125]

#Biomedicalandbiololicalscience , #Olivercarvajalgômez has an extremely legitimate question. Would you be so kind as to oblige with an answer? Kary Mullis, the inventor of the PCR test had specifically stated it is an *AMPLIFICATION* test only. No different than turning the volume up on a stereo so loud the *amplifier* would blow out a speaker as would the thermal heat.

[@baghiballsakh82]

I found these two patents from Kary Mullis and his team for PCR. Might help 🤷 System for automated performance of the polymerase chain reaction (US Patent US5656493A) This method is especially useful for performing clinical tests on the DNA or RNA from a fetus or other donor where large amounts of the DNA or RNA are not readily available and more DNA or RNA must be manufactured to have a sufficient amount to perform tests. The presence of diseases which have unique DNA or RNA signatures can be detected by amplifying a nucleic acid sample from a patient and using various probe procedures to assay for the presence of the nucleic acid sequence being detected in the test. (Patent number: 4,965,188) Various infectious diseases can be diagnosed by the presence in clinical samples of specific DNA sequences characteristic of the causative microorganism. These include bacteria, such as Salmonella, Chlamydia, Neis seria., viruses, such as the hepatitis viruses, and parasites, such as the Plasmodium responsible for malaria.

[@atpsmpss4201]

From Spain I want to congratulate you for the wonderful videos that you do. Your explanations are simple and at the same time very complete. Thank you so much for your work

[@mpfmax0]

You have the annealing temperature limits backwards: too high means no binding and too low means unspecific binding. Also you have the reverse and forward primer backwards as well on your graphic.

[@paolagissellepicazoflores7760]

thank u random human being on the internet for explaning this before my final genetics exam (second semester med student), i hope your charger works from all angles

[@desalegnbelaytakele6267]

how was the result of your exam then?

[@kasahunfirew3592]

🔥❤️

[@michaeltolkachov4749]

actually, the higher the temperature of the primer annealing the higher the specificity. This is why when doing a gradient PCR, we typically take the products from the highest annealing temperature, it is also a good way to test newly synthesized primers and save time :) The denaturation time and temperature often depends on the buffer composition and used polymerase (e.g. Kapa2G has 94 and Q5 of NEB has 98 recommended) though usually, it works anyways. I'm mentioning just in case, cool video!

[@anniesaminaiken1665]

Hi sorry I'm really confused @ 12.25 you mentioned that if the annealing temperature is too low, the primer will not bind and if the temperature is too high the primer will bind non-specifically. But I was taught the opposite and no matter how much I research into it, I find that I'm correct. if the temperature is too high, there will be no hydrogen bond formation and the primer will remain dissociate, and if the temperature is too low the primer will bond non-specifically.

[@priyankamanuja1617]

U r correct ..may be it's by mistakenly said

[@mralamtech1724]

Hello Annie

[@zifantang3792]

Thanks for correction

[@AndrewCharnley]

Looks like you were probably correct. See the commentary from: mpfmax0 7 months ago "...You have the annealing temperature limits backwards: too high means no binding and too low means unspecific binding. Also you have the reverse and forward primer backwards as well on your graphic..."

[@ehydrianluwi]

you are correct, high temperature denatures the hydrogen bonds thus breaking the bond.

[@pramitanindyasaraswati9308]

Well-explained! I get a lot of benefits by watching this 20:34-minute video, I really appreciate it, thank you for sharing and looking forward to watching other videos from your channel :)

[@calarzt3453]

Big Thanks for you 🌷 My specialist genetics and biotechnology.. Please video about COVID_19 PCR ..and technical procedure And which the perfect pcr device to diagnosis this infection

[@estellejames9993]

Thank you for instruction on a complex scientific procedure and making it understandable for a new student.

[@BlackZero880]

Dear mam, i just love you way of explanation, i am preparing for the interview and helping me a lot. Keep posting such valuable lecturres. Thank you very much, do you have a video on RT PCR? plz reply

[@fanyang6608]

Thank you for your wonderful explanation. I still have one question: When the elongation step starts, the dNTPs begin to bond the specific DNA sequences after the primer. But how to stop them at the terminal of the specific sequences. There is no such things like primer to stop them at the termination.

[@caroed2768]

the first two newly synthesised chains won't have the right length, they'll be longer, but after that you'll start having single strands of the right length. The first time you'll have double stranded dna of the right length (on both strands) will be cycle 3, and from there the number grows exponentially. This is clearly shown in this video kzbin.info/www/bejne/n4LWpmaBr56Bj9E (minute 2.00)

[@fanyang6608]

@@caroed2768 Thank you! My friend, you answered the question that had been bothering me for half a year!

[@fanyang6608]

@@aburahat4653 Thank you for your answer. I have understood the process now

[@Belleville197]

The best PCR explanation video I've seen so far. Good job girl.

[@minetopcuoglu266]

Thank you. This a clear and easy explanation. You have done a better job than my lecturers in medical school.

[@alcepeda6843]

Highly appreciate your video, can you please add a note in the description with a correction regarding the primer binding temperature tolerances that are mentioned at 12:25. The details are in Annie Jay's comment below. You have a great gift for teaching as is obvious in this video. Your graphics are highly effective.

[@terrie001]

Thanks! Question - how does the DNA Polymerase knows when to stop the elongation step (I know it starts at the primer, but how about the end)? I guess the DNA is super long, so if my target is only 1000 base pairs, and I don't need 5000 base pairs DNA strand.

[@hendrawanbakri9363]

Terrie 000 it's about the time, basically 1 minute is time that needed to copy about 1000 bp, when you reached that limit then decrease or increase the temperature so the dna polymerase wont work properly

[@emmieali6397]

انا عربية وفهمت عليكي لغتك جدا بسيطه وسهلة. Thank u

[@jimbean4751]

ty for this informative and educational video Especially now it's important to know how this works.

[@fatunbarinoladayo5699]

Thanks for the detailed method of teaching..😊😊😊

[@NicoBertix]

Very well explained, thanks for this...The way you say bye at the end is so cute :-)

[@bryanjones7126]

Outstanding overview! Well done!

[@zehraishaq4955]

I don't understand people who dislike this video ,,what is their problem.

[@luciachp3519]

Thank you very much, I loved the explanation

[@yun-ching-sun]

Thanks for the clear explanation. Let me know the details of PCR or how it works.

[@AA-gl1dr]

i could listen to you teach for hours on end. tremendously easy to understand.

[@ssierrat]

Thanks a lot for your videos, they are very helpful, you have a special capacity to explain complex subjects in a simple way. Thank you.

[@aliyas8733]

Wow thank you very much you’re lifesaver

[@hankschrader2353]

The annotation of the primers is wrong! The reverse primer binds to the template. Its also just the orientation in which the polymerase then binds and then extends the primer in relation to the direction of the sequence.

[@drzenaAcademy]

شرح جميل لكن التئتئة متعبة للسامع

[@esrakayikci934]

Hi, could you tell me the references maybe?

[@abdulazizadamhassan6252]

What is exponential phase and non exponential phase

[@pritichristian2673]

Thanks. Great information

[@vincentmusilikani6717]

Thank you very much🙏🏾

[@shailashermintania4007]

Very nice & clear presentation

[@adnan.q.753]

This was explained really nicely.

[@karenaw3428]

Thank you so much for this video! This was so awesome and helpful!

[@biomedicalandbiologicalsci4989]

Thank you for the nice comment :)

[@nadiasaad702]

Thanks for the amazing video Your explanation is so perfect I will follow your videos in the future Thx

[@Wyvernnnn]

Does this type of amplification yield more mutations than how would naturally occur in the human body?

[@mervatmorad5052]

thank you, it is simple and helpful

[@esrakayikci934]

very well explained! thank you for sharing..

[@usoewynn4705]

Thanks . Excellent explanation. Also for RT. PCR.

[@poornimasirvaiya7949]

Thank you so much mam 🙏🙏 this video cleared all my doubts 🤩 really very very helpful. 🙏🙏

[@martinmazukiewicz]

Hello everybody. Question : i understand why the polymerase starts copying the strand (because it's the place where the primer is, with its OH), but i don't explain why it's ending the copy at the end of the chosen sequence. Noboody has told him. Why it's not going further ? There is nothing to stop it ? Thanks for answering. ;-)

[@martinmazukiewicz]

Ok, i found the answer. Easy when you know.... For those who don't know, here is the explanation : www.sciencebuddies.org/science-fair-projects/ask-an-expert/viewtopic.php?t=987

[@marwagouda6819]

It is a very interesting way to deliver knowledge. Much thanks

[@deegallarupananda6927]

Thank you very much!

[@BlackZero880]

I just Love ur way of explanation mam....from india

[@luisatali]

Does the taq polymerase just stop at the end of the desired dna length (so at the end of the red marked dna in the video) or does it synthesize the rest as well?

[@kedirmohammed1623]

I apreciat you

[@animeshsamanta4503]

Very beautiful explanation... Thank you very much

[@sanjeevkumarnitrourkela6637]

Hello Ma'am...very nice explaination..

[@terrie001]

In the lab, how much is "one copy" of DNA? Like 10 uL of DNA sample extraction, 100 uL? Or how much do people generally start out with, you know?

[@funnyanimalshorts643]

17:54 Yes but it is literally exponential growth.

[@exophthalmos1]

..and she literally gave an example with an exponent. What is your point?

[@funnyanimalshorts643]

@@exophthalmos1 Exponential growth: Exponential growth is a specific way that a quantity may increase over time. It occurs when the instantaneous rate of change of a quantity with respect to time is proportional to the quantity itself. This also may help kzbin.info/www/bejne/bWLcpntpa5qFq8k

[@alwaysontherise1]

Beautifully done! Very clear explanation

[@KhaLed-pb4pu]

what is the difference between Parent DNA and DNA contents ?

[@صاحبعباسالكبيسي]

Thank you very much for this explanation ❤

[@fatimahalnasser12]

well done !! very clear explanation.

[@hayaalmly6083]

Thank you!!! you made it really easy and simple!

[@Abdulhadi-vn7ui]

thank you so much

[@nuwanthachamikara5823]

Thank you. Very useful content 👌

[@amayamgoyeng]

I see the light ~~~~

[@nikolinakrysteva7770]

Thank you for very interesting and nice presentation of PCR. I will used to offer to my studenrs to see and to enjoy the news in the practise biochemistry. Wish you all the best and to show new methods and success in biochemistry. d-r Krusteva

[@alitalib6342]

Hi. How can I get this presentation please?

[@francescabaysa1773]

THANK YOU!

[@FlashMusic2U]

Great Video

[@نمرودنمرود-خ5ل]

Thanks 👍👍👍

[@SaiKumar-wu3ge]

Exalent mam thank for this vedio

[@jawharahahmed5031]

thank you so much really you made it easy

[@shauryadumka9865]

U explained it well.. great work..

[@DrBill-wd1lu]

so good👍👍

[@tianyusong7735]

Best!!! Thank you!!

[@bereketwake6622]

It's interesting...thank you!!!

[@critthepoet9160]

Very cool

[@queenieferolino5061]

i really don't have a backgrount in pcr and we will have a defense abt this thanks for the infosss

[@biomedicalandbiologicalsci4989]

meanyoongi 0309 thank you for the comment... stay tuned ... and suggest the video to your colleagues ;)

[@biomedicalandbiologicalsci4989]

Welcome ... stay tuned :)

[@kamranhussain5602]

nice video

[@isuriupadhya535]

Great

[@surendraangom9743]

Nicely explained. Thank you.

[@ezendianefojosiemkparu1951]

TX MUM, U ARE EXCELLENCE PERSONIFIED

[@anshisview7280]

reallly good......itz help me alot...suprb explanation

[@aktv2137]

Thank you very much . i watched it from Bangladesh. this video is helpful for us.

[@maisasantos2654]

Thank you so much! I have a test and this video helped me a lot!!

[@biomedicalandbiologicalsci4989]

I am glade to hear that :)

[@quratulainnuser6076]

thank you very much its superb

[@biomedicalandbiologicalsci4989]

You are welcome :)

[@amnaijaz7384]

Great work. But i have a question that how do we quantify viral load as in HCV through PCR?

[@CarlosSantos-so5ww]

In order to quantify any viral load, you don't use PCR. You need to use qPCR.

[@tzatzikosouvlaki]

Could you do a video about 16s RNA sequencing ? about the 2-step PCR and how the elongation of the adapters is done. Perfect job , keep on :) :)

[@noorpk]

We can discuss about adopters.

[@tzatzikosouvlaki]

@@noorpk ok i m waiting for your next one

[@mohammedibrahim4500]

Thank you .You are special

[@rukhsorasultonova]

thank you well done

[@2morganj]

very good information, thank you...

[@docshaheen6741]

Excellent lecturer

[@sabbirahmed8602]

wow, your explainassion is so easy. thank you

[@andrewmcgee1351]

Great explanation!

[@sarahe.s7231]

Thank you very much ❤️

[@anleyteferra7846]

Thank you!

[@zeroframe5415]

great video! thank you

[@zargairostamkhill3926]

that was great to learn more about pcr thanks again

[@mohameda.hassany6562]

you are excellent and amazing

[@firas4199]

well done

[@llxu1685]

Thank you!!! It's helpful for me!!!

[@rimel6446]

Perfect

[@mohamedsamir6751]

Thank you

[@rezenehabte434]

WELL PRESENTED

[@alizahranmohd4095]

Asante sana.

[@vinodvenugopal5143]

thank you, explained well