Polymerase Chain Reaction (PCR) Protocol

  Рет қаралды 64,564

Addgene

Addgene

Күн бұрын

Do you have a short region of a DNA molecule that needs to be copied? Alyssa, a QC Scientist at Addgene, walks you through the PCR process. For the full protocol text, visit www.addgene.org/protocols/pcr/
A standard Polymerase Chain Reaction (PCR) is an in vitro method that allows a single, short region of a DNA molecule (single gene perhaps) to be copied multiple times by Taq Polymerase. From a single copy of DNA (the template), a researcher can create thousands of identical copies using a simple set of reagents and a basic heating and cooling (denaturing and annealing) cycle. The process became automated with the discovery of a heat resistant DNA polymerase from the thermophilic bacterium, Thermus aquaticus (Taq). Taq polymerase can withstand many heating and cooling cycles, which would denature DNA polymerases from other species.
___________________________________________________________________________
0:00 Intro & Overview
1:35 Materials
2:29 Getting Started in the Lab
3:20 PCR Process
5:01 Agarose Gel Prep & Downtime
5:15 Running Your Gel
5:30 Troubleshooting
___________________________________________________________________________
Credits
Featuring: Alyssa Cecchetelli
Directed, Animated, Edited, & Written by: Quintin Marcelino
Music by: Lauren Duski, Vibe Mountain, and RKVC, courtesy of KZfaq Audio Library

Пікірлер: 38
@addgene
@addgene Жыл бұрын
Addgene does not regularly monitor comments posted here, so we may not see your question immediately. We’d be happy to answer any questions sent to help@addgene.org as soon as possible. Please include the name of the video along with any questions so our support team can help. Thanks!
@asmaelaouina7047
@asmaelaouina7047 2 жыл бұрын
Those funny moments are amazing... great job guys keep up the good work
@amankaur9969
@amankaur9969 Жыл бұрын
I have attained better understanding of elution and PCR after watching your video. Thank you so much.
@roshnibhatt2801
@roshnibhatt2801 Жыл бұрын
All protocols are given complete information, Thankyou
@nermeenabdelmoez6347
@nermeenabdelmoez6347 3 жыл бұрын
Brilliant! Thank you so much!❤
@hemastandard7649
@hemastandard7649 3 жыл бұрын
Amazing presentation
@jasonlafontaine9882
@jasonlafontaine9882 2 жыл бұрын
Absolutely fantastic
@ricardoguta290
@ricardoguta290 2 жыл бұрын
THANK YOU VERY MUCH IT WAS VERY HELPFUL VIDEO
@jacquelinevaliente1209
@jacquelinevaliente1209 3 жыл бұрын
Superb!
@hidayatullah6740
@hidayatullah6740 4 жыл бұрын
Great maam!...
@sabah.bsabah.b1559
@sabah.bsabah.b1559 11 ай бұрын
thank you
@liyaaudinah5718
@liyaaudinah5718 10 ай бұрын
Thanks❤
@wferrand
@wferrand 9 ай бұрын
Very cool
@notabigdeal7785
@notabigdeal7785 3 жыл бұрын
Overall very helpful video I appreciate the time and effort you put into explaining the stages of annealing and denaturing-- very helpful. Some more details on the ingredients list, especially explaining mM and buffers, and how they apply to the overall chemistry of PCR, would be helpful, thank you so much!
@addgene
@addgene 3 жыл бұрын
Thanks for the feedback! We try to pack as much info into videos as possible, but sometimes we can't cover everything. But we do have a written PCR protocol that digs into some more details: www.addgene.org/protocols/pcr/
@kano1957
@kano1957 4 жыл бұрын
Nice
@ranjanbiology
@ranjanbiology 2 жыл бұрын
Lovely
@Stronger.119
@Stronger.119 2 жыл бұрын
❤️I love addgene's website
@europhile2658
@europhile2658 3 жыл бұрын
very nice video. I like the solutions to the problems
@luckyrook1246
@luckyrook1246 4 жыл бұрын
PCR baby!! WOOHOOOO!!!!!
@ermiyasshibesh3396
@ermiyasshibesh3396 8 ай бұрын
Very constructive videos! Please I need to get PCR-RFLP steeps?
@user-vf8bj3pn1t
@user-vf8bj3pn1t 9 ай бұрын
Plz share info about genomics .. Proteomics... Transcriptomics...
@stephenolinga9236
@stephenolinga9236 Ай бұрын
V good
@Best_Play1996
@Best_Play1996 3 жыл бұрын
Really wonderful video 😍😍 can you explain me in detail about threshold value ,limitation of pcr ...
@addgene
@addgene 3 жыл бұрын
It looks like you're talking about qPCR? This guide has some more information on the cycle threshold for qPCR www.thermofisher.com/us/en/home/life-science/pcr/real-time-pcr/real-time-pcr-learning-center/real-time-pcr-basics/real-time-pcr-understanding-ct.html
@user-vg8tq9zi4j
@user-vg8tq9zi4j 11 ай бұрын
Why we but the componant on ice at the beginning?
@nabilahshudirman7000
@nabilahshudirman7000 4 жыл бұрын
Love
@aswinipriya7558
@aswinipriya7558 3 жыл бұрын
Really amazing, I have doubt , if target is amplified remaining PCR components where it goes, another one doubt during extension , how it sharply terminate at target site - please explain
@christeredvardsson8078
@christeredvardsson8078 2 жыл бұрын
All want in life is the PCR machines to be covered with cloth that can dramatically be removed with a soundbite cheering me on during setup.
@ammarsohail7901
@ammarsohail7901 3 жыл бұрын
Content were well arrange but less details kindly add some more details our all👍👍❤️
@addgene
@addgene 3 жыл бұрын
Thanks for the feedback! We try to pack each video with as much information as possible, but some things just can't make it into the video. For a more in-depth look, you can check out our written protocol here: www.addgene.org/protocols/pcr/
@hosseinvasighi4279
@hosseinvasighi4279 Жыл бұрын
Tanx
@toxicmooyahperson1847
@toxicmooyahperson1847 2 ай бұрын
good ass video
@lenilestari8908
@lenilestari8908 3 жыл бұрын
Anyone know this method is conventional or real time PCR?
@addgene
@addgene 3 жыл бұрын
Hello! It's conventional (end point) PCR.
@Therealtalkpodcast1115
@Therealtalkpodcast1115 2 жыл бұрын
Why not add mgcl2 initially?
@addgene
@addgene 2 жыл бұрын
Thanks for the question. Magnesium chloride is already included in the 10X Taq buffer, but some PCR reactions may require more magnesium chloride to improve DNA amplification. Therefore, adding more magnesium chloride to a PCR reaction that is not working might improve your results. However, excess magnesium chloride can decrease the binding specificity of your primers, so we suggest trying a PCR reaction under standard conditions first.
@strawberry_nya
@strawberry_nya Жыл бұрын
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