Polymerase Chain Reaction (PCR) Protocol
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- čas přidán 30. 06. 2024
- 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.
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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
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Credits
Featuring: Alyssa Cecchetelli
Directed, Animated, Edited, & Written by: Quintin Marcelino
Music by: Lauren Duski, Vibe Mountain, and RKVC, courtesy of CZcams Audio Library - Věda a technologie
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!
Those funny moments are amazing... great job guys keep up the good work
I have attained better understanding of elution and PCR after watching your video. Thank you so much.
All protocols are given complete information, Thankyou
Brilliant! Thank you so much!❤
Amazing presentation
Absolutely fantastic
THANK YOU VERY MUCH IT WAS VERY HELPFUL VIDEO
Superb!
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!
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/
Great maam!...
thank you
Thanks❤
Very cool
❤️I love addgene's website
Nice
Lovely
very nice video. I like the solutions to the problems
PCR baby!! WOOHOOOO!!!!!
Very constructive videos! Please I need to get PCR-RFLP steeps?
Really wonderful video 😍😍
can you explain me in detail about threshold value ,limitation of pcr ...
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
Plz share info about genomics .. Proteomics... Transcriptomics...
Why we but the componant on ice at the beginning?
V good
Love
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
Content were well arrange but less details kindly add some more details our all👍👍❤️
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/
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.
Tanx
Anyone know this method is conventional or real time PCR?
Hello! It's conventional (end point) PCR.
good ass video
Why not add mgcl2 initially?
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.
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