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...
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
*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.
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.
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
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 :)
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!
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
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.
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?
#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.
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.
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
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.
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..."
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?
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
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.
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
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.
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 ru-vid.com/video/%D0%B2%D0%B8%D0%B4%D0%B5%D0%BE-iQsu3Kz9NYo.html (minute 2.00)
