PCR (polymerase chain reaction) in detail

This pcr lecture explains polymerase chain reaction in details.
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The polymerase chain reaction (PCR) is a biochemical technology in molecular biology to amplify a single or a few copies of a piece of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence.
Developed in 1983 by Kary Mullis,[1][2] PCR is now a common and often indispensable technique used in medical and biological research labs for a variety of applications.[3][4] These include DNA cloning for sequencing, DNA-based phylogeny, or functional analysis of genes; the diagnosis of hereditary diseases; the identification of genetic fingerprints (used in forensic sciences and paternity testing); and the detection and diagnosis of infectious diseases. In 1993, Mullis was awarded the Nobel Prize in Chemistry along with Michael Smith for his work on PCR.[5]
The method relies on thermal cycling, consisting of cycles of repeated heating and cooling of the reaction for DNA melting and enzymatic replication of the DNA. Primers (short DNA fragments) containing sequences complementary to the target region along with a DNA polymerase (after which the method is named) are key components to enable selective and repeated amplification. As PCR progresses, the DNA generated is itself used as a template for replication, setting in motion a chain reaction in which the DNA template is exponentially amplified. PCR can be extensively modified to perform a wide array of genetic manipulations.
Almost all PCR applications employ a heat-stable DNA polymerase, such as Taq polymerase, an enzyme originally isolated from the bacterium Thermus aquaticus. This DNA polymerase enzymatically assembles a new DNA strand from DNA building-blocks, the nucleotides, by using single-stranded DNA as a template and DNA oligonucleotides (also called DNA primers), which are required for initiation of DNA synthesis. The vast majority of PCR methods use thermal cycling, i.e., alternately heating and cooling the PCR sample through a defined series of temperature steps. In the first step, the two strands of the DNA double helix are physically separated at a high temperature in a process called DNA melting. In the second step, the temperature is lowered and the two DNA strands become templates for DNA polymerase to selectively amplify the target DNA. The selectivity of PCR results from the use of primers that are complementary to the DNA region targeted for amplification under specific thermal cycling conditions. Source of the article published in description is Wikipedia. I am sharing their material. Copyright by original content developers of Wikipedia.
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[@leorix123]

Basically passed all my exams from organic chem to molecular biology related subjects in uni by refreshing the contents in my notes through ur lectures. Keep it up sir! Well done!

[@h.upekkhaabhayarathne2274]

We do watch your videos all the time and they are so much helpful for us in biotechnology sir. We do almost all the exams in the university by watching your videos. You are capable of teaching even the hardest parts, in simple ways. We like to watch videos which you yourself teach in the white board, very much. Thank you sir...bohoth bohoth shukriya!!

[@shomusbiologyofficial]

Thank you so much for appreciating my efforts

[@chocalateicecream]

I wanted this a few times- thank you for breaking down the process of PCR. You don't waste time on unnecessary words or pauses like many "youtube teachers" do,

[@ThiNKBiologyThiNKVISION]

The Best and Complete PCR Explanation Video Ever...👏👏
There are 100s of Videos on PCR, But this the best...!
Thank You...!🙏🙏

[@shomusbiologyofficial]

Thank you so much for appreciating my efforts. Glad to hear that you're getting benefit from my lectures

[@alashadbin3814]

thanks for perfect videos you share they're both informative and productive. It's brilliant that we have such awesome lecturers like you in TH-cam who are trying their best to inform others

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Thank you so much for appreciating my efforts

[@anamikahalder2102]

God bless u sir . Ur doing a grt help to thousands of people with ur beautiful videos..high quality teaching ♥️

[@shomusbiologyofficial]

You're welcome. Glad to hear that you're getting benefit from my lectures

[@architadas4604]

BEST PCR LECTURE EVER🙏thanks a lot sir

[@shomusbiologyofficial]

You're welcome

[@Ramya.85]

i am getting hope to clear exam only because of your videos. after 8 yrs gap, i started to persue my carrier, its such a long break , ur videos given me much confidence . i hats off u. u r my role model thanks shomu sir.

[@newthinker5829]

Hi r u working now... I am also started studying after 8 yrs of gap hoping to get a job soon

[@annajoy1948]

your videos are good for brushing up concepts rather than reading all the books again when time is short. thanks.

[@haryobolodewo4669]

I am very blessed found your tutorial lecture on Molecular Biology in the you tube, you gave a simple way how to understand the application and knowledge about molecular biology, you are very kind to share your knowledge to everybody in the world.....hope you get present from the God....salam from Indonesia....

[@wenjinghe9629]

IT IS SO HELPFUL!!! your explanation is so much better than my tutor's. thank you so much

[@Anabelle6493]

a really complete and informative video. good job and thank you :)

[@hajrabibi8773]

Very interesting sir as you said .thank you very much for the lecture.you have provided us with very minute details which are Essential for attending the long questions ans . ❤️❤️

[@mohdkaleemkhan9994]

Nice slide and explained every step in authentic way...

[@vinitasharma4578]

U ARE THE BEST.... HELPING ME ALOT DURING MY STUDIES. THANKS

[@terrybash5194]

Thank you so much DR RORPOPOR HERBAL on TH-cam you saved my life from this deadly PCR virus, I got cured within 14daysly herpesly herpes..........

[@bansisharma720]

hi shomu i have a question reagarding pcr reaction.. why initial pcr product produce long and varying size product in reaction, and fina reaction produced shorter and uniform size product?

[@afiqah443]

very helpful . The content is very detailed. Great job sir.

[@shomusbiologyofficial]

Glad to hear that you are getting benefit from the videos

[@sukanyapaul8666]

It is so much helpful, very informative and descriptive video. Thank you sir😇

[@shomusbiologyofficial]

Glad to hear that you're getting benefit from my lectures

[@yumnafouzer5923]

i'm starting to have hope, Thank you sir

[@shomusbiologyofficial]

You're welcome

[@marisettylokesh2620]

Tq so much for better understandingsir......I like ur teaching skills sir.....

[@shomusbiologyofficial]

Glad to hear that you're getting benefit from my lectures

[@biniamtesfazghi8181]

it is very helpful my lecturer recommend me to watch this

[@shomusbiologyofficial]

Glad it helped

[@সাদমোহাম্মদ]

আপনার video গুলো অনেক সুন্দর। অনেক concept clear হয়েছে। আপনাকে অনেক ধন্যবাদ।

[@shomusbiologyofficial]

Thank you for appreciating my efforts

[@andreainzaghi7373]

thank you Dr Shomu

[@shomusbiologyofficial]

Glad to hear that you're getting benefit from my lectures

[@jashodaksuthar8934]

Got one question! when a gene is encoded on a single strand of DNA called template strand then why do we perform PCR on both strands of DNA using 2 primers?

[@rajithaanand9879]

Thank you sir for the good work .... I have understand the concept of PCR very easily through ur lecture, it's very helpful. . Please upload more videos related to this . Your explanation is very good ....

[@totorocute9638]

This is very helpful, my doctor ask us to watch your video and it best decisions ever,

[@shomusbiologyofficial]

Thank you so much for appreciating my efforts

[@terrybash5194]

Thank you so much DR RORPOPOR HERBAL on TH-cam you saved my life from this deadly PCR virus, I got cured within 14daysly herpesly herpes..........

[@iqraijaz5476]

Thanks alot for making this subject easier may God bless you

[@misbahnaik4987]

I didn't understand this process even though my teacher took 3 hrs to explain it.. Thanks sir for the explanation

[@medschoolradio]

Thanks...you just made everything so clear!!

[@priyatalib9330]

Your all videos are really helpful. Thank you so much.

[@arpanbanerji9962]

Thank you very much sir...oshonkhyo dhonnobad

[@shomusbiologyofficial]

Thank you. Glad you liked my lectures

[@avnigoel5420]

Hello sir, your videos are very helpful to all of us ( me and my batchmates )sir we all just watch your videos and we all have passed our exams with good Marks thank u soo much sir for making every topic so simple to understand. sir I am facing problem in plateau effect in PCR ,I am not able to understand it, so sir can you please help me out

[@shomusbiologyofficial]

Thank you. Glad you liked my lectures. Please subscribe and share

[@sanjibkalita403]

very very lucid explanation.
Thank you sir.

[@yousufbaloch5140]

How sir the dna denaturate at specific site, if you provide heat it should denaturante completely?

[@Easybotanytastybotany]

Very helpful thank you so much🙏🙏🙏

[@shomusbiologyofficial]

You're welcome

[@aparnaaparna9992]

Sir I one big request.. can you make one video about enterococcus feacalis full detail

[@jaserahsyed441]

A primer may bind anywhere on the DNA, obviously to ours complementary sequences but it's not always necessary that these sequences will be present at the very start. So the sequences where primer doens't bind how are those sequences amplified?

[@sathimerugu821]

sir, i am confusing the order of the videos, recombinate DNA Technology 61 tutorials are there , but which one is 1st , 2nd..........61 videos, this is my confusing , i hope you pls clearify , thank u.

[@microscopy3852]

very good technique

[@vidhisambhvani8276]

hello tysm for this video.u r genius.u helped me in molecular biology n genetic engineering so much..can u plz provide me yr written material or books...???

[@khadijadabeer6284]

inspite of being such a good method of amplification why cloning is not completely replaced by pcr?

[@maalyahmed]

Thank you very much, very informative and helpful video

[@sobiasana8365]

Sir why primase is not used ?is that bcz of small size of DNA fragment that is to be copied

[@demissedakone2980]

Great lecturer, thank you

[@Encrypted628]

Thank you very much for your video lecture, much appreciated.

[@doveofpeace9120]

Thank you very much just a question about the oligonucléotides they are of DNA or RNA????

[@riyabarik7537]

Very informative video. hoping for t same for other techniques....thanks slot it really helped me

[@dishapatel2911]

sir plz explain chemical. synthesis of dna & its 3 types

[@savitact7960]

Thankyou very much for such an informative video.. keep it up.

[@shomusbiologyofficial]

+Savita Ct thank you. Glad you liked my lectures

[@anshulrana5614]

Thank you so much sir 🥰🥰🥰🥰

[@shomusbiologyofficial]

You're welcome

[@Shwetachaturvedi25]

Why all videos on PCR r not available at u tube???plz make them easily access for all students those who can't afford coaching...

[@sriparnadas7781]

Really very helpful sir....plse sir can u explain suicide pcr in another tutorial video.

[@maheesiriwardhana1208]

Thank you very much sir.

[@shomusbiologyofficial]

You're welcome

[@terrybash5194]

Thank you so much DR RORPOPOR HERBAL on TH-cam you saved my life from this deadly PCR virus, I got cured within 14daysly herpesly herpes..........

[@fatirafimor1326]

Thank u very much sir. This really help me to understand more about PCR.

[@Noor-ty1xt]

Why both primers ?

[@genetukassahun2679]

good lecture

[@shomusbiologyofficial]

You're welcome

[@dikshatiwari2562]

What are the different enzymes used for this process

[@vijaybhaskarreddykoyle7347]

hello sir, help me to clear my doubt that initially two primers are used for 1st cycle but how many primers are used for 30
cycles

[@bhanu549]

thanks a lot...very nicely explained

[@shafiullahshehu4050]

Work well done. thanks sir

[@Attamatic]

impressive command of English

[@romi6374]

In which base pair does the primer starts binding in annealing steps? It binds to complementary sequence but in which exact bp does it bind?

[@craftsandscience860]

It binds first at the 3' end of the primer. That's why the 5 base pairs on the 3' end of the primer are very important In order to promote a proper annealing to the DNA complementary strand. Therefore, better to design a primer with G or C nucleotide at the very end of the 3' end of the primer because it has a strong binging efficiency

[@healthsupport5218]

Hi I have a very basic Question regarding PCR. PCR is used to amplify content of DNA. Say for ex. My sample contains 38ng/microL concentration of DNA, using PCR Am I making my solution more concentrated? (from 38 ng to 38 micro/mg?) Please clarify..

[@csmobilesolutions8553]

Health Support yes

[@mikehunt3426]

I was wondering... What would happen if the two primers were on the same strands ?? Would there be any PCR's product or nothing ?

[@waiting12]

Mike Hunt thinking same ...if we suceeded to any how artificially ....dna manuplating by pahogens could be reduced...

[@zhsaaad]

Thank you so much. It was very helpful.

[@himanigoel624]

it was an awesome video. 😊 very helpful

[@David_Wigren]

Excellent video!

[@Nikkishoko]

Thank you very much! This is really helpful!

[@sabaismail8622]

Does anyone have any experience with trying to PCR with loading dye present? Or any ideas on how to get rid of it so that it doesn't inhibit the enzyme?

[@samthakur4038]

excellent video

[@sagnikbiswas7045]

we need to amplify only one strand of the DNA,as,the sense strand contains the required fragment of DNA,then why do we need the anti-sense strand of DNA for amplification??? why do we add two primers in that case,,,,when we need to amplify only the sense DNA strand?

[@SuperAnnanymous]

Sagnik Biswas Because anti sense strand is complementary to the sense strand. We actually get sense strand by amplifying an anti sense strand. This is my thinking. I maybe wrong.

[@sagnikbiswas7045]

You're right ☺

[@firozsheikh3352]

great job God bless you 🙌🙋

[@amisheriff709]

very informative

[@officialperson8802]

Thank you bhai 😭

[@shomusbiologyofficial]

You're welcome

[@AD-dy1hi]

Heat labile means heat sensitive. So Taq polymerase is not heat sensitive and neither heat labile!!!!
Thank you for your video.

[@shomusbiologyofficial]

You're welcome

[@mezbauddinrobin6167]

Helpful..

[@shomusbiologyofficial]

You're welcome

[@medschoolradio]

Very nice...

[@sweetysharma5268]

very informative video

[@shomusbiologyofficial]

Thank you. Glad you like my lectures. Stay tuned.

[@sonailkhattak8180]

what is means by melting temprature

[@analudmilamedeiros6636]

very good.

[@viduradias4646]

Thank You!!!

[@shomusbiologyofficial]

You're welcome

[@ritikasingh8809]

Thanks

[@aqsaali2774]

Wel Done sir !

[@kingcorneliusnoamesi6118]

great video. tnx

[@razuahmed7736]

IT IS VERY MUCH HELPFULL

[@sabitasinku9548]

Sir iss topic ki note kaise download karein

[@shomusbiologyofficial]

Don't have note now

[@SUNILGBA]

very good keep it up

[@Jacksonjeans1]

thank you

[@nameirakpambidyanandasingh9534]

it sounds good thanks alot

[@akriti6152]

what taq polymerase is important

[@mdwazid1548]

akriti nipu coz it can resist high degree of temperature

[@morsmb1]

ty very much

[@pritidesai1924]

nice

[@pakhisingh6378]

Grt👍

[@shomusbiologyofficial]

Thank you

[@yourmajesty1361]

I am afraid that this video is 30 min. hope i will get it

[@mbitasumbwe3169]

PCR made easier thanks

[@fatimabintezahir3956]

👏👏

[@وردةالحياة-ق8ح]

اكو عرب اهنا 😂😂

[@maczampieri]

nice english accent!!!

[@kundanyadav9042]

Plz speak a little faster