Hot start PCR
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11 سال پیش
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This PCR series lecture explains
This PCR series lecture explains the hot start PCR prionciple and use in gene amplification. http://www.shomusbiology.com/
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The hot start PCR is a modified form of Polymerase chain reaction (PCR) which avoids a non-specific amplification of DNA by inactivating the taq polymerase at lower temperature. In this dsDNA is denatured by heating the sample at its denaturing temperature and then the temperature is suddenly reduced to 55 degree C at which primer and Taq-polymerase is added, but here the difference arises i.e. specific antibodies are used to block this Taq-polymerase at annealing temperature. Now when the temperature raises for amplification to 72 degrees, the specific antibody detaches from Taq-polymerase and the amplification with greater specificity starts. In conventional PCR, the Taq DNA polymerase is active at room temperature and to a lesser degree, even on ice. In some instances, when all the reaction components are put together, nonspecific primer annealing can occur due to these low temperatures. This nonspecific annealed primer can then be extended by the Taq DNA polymerase, generating nonspecific products and lowering product yields. Hot Start PCR significantly reduces nonspecific priming, the formation of primer dimers, and often, increases product yields[citation needed]. In Hot Start Long and Accurate PCR, the impact on yield can be dramatic[citation needed]. Classic methods, while effective, involve additional handling and increased risk of contamination. [1] Source of the article published in description is Wikipedia. I am sharing their material. Copyright by original content developers of Wikipedia.
Link- http://en.wikipedia.org/wiki/Main_Page
Download the study materials here-
http://shomusbiology.com/bio-material...
Remember Shomu’s Biology is created to spread the knowledge of life science and biology by sharing all this free biology lectures video and animation presented by Suman Bhattacharjee in YouTube. All these tutorials are brought to you for free. Please subscribe to our channel so that we can grow together. You can check for any of the following services from Shomu’s Biology-
Buy Shomu’s Biology lecture DVD set- www.shomusbiology.com/dvd-store
Shomu’s Biology assignment services – www.shomusbiology.com/assignment -help
Join Online coaching for CSIR NET exam – www.shomusbiology.com/net-coaching
We are social. Find us on different sites here-
Our Website – www.shomusbiology.com
Facebook page- Facebook: ShomusBiology
Twitter - Twitter: shomusbiology
SlideShare- www.slideshare.net/shomusbiology
Google plus- https://plus.google.com/1136485849827...
LinkedIn - LinkedIn: suman-bhattacharjee-2a051661
Youtube- thefunsuman
Thank you for watching
The hot start PCR is a modified form of Polymerase chain reaction (PCR) which avoids a non-specific amplification of DNA by inactivating the taq polymerase at lower temperature. In this dsDNA is denatured by heating the sample at its denaturing temperature and then the temperature is suddenly reduced to 55 degree C at which primer and Taq-polymerase is added, but here the difference arises i.e. specific antibodies are used to block this Taq-polymerase at annealing temperature. Now when the temperature raises for amplification to 72 degrees, the specific antibody detaches from Taq-polymerase and the amplification with greater specificity starts. In conventional PCR, the Taq DNA polymerase is active at room temperature and to a lesser degree, even on ice. In some instances, when all the reaction components are put together, nonspecific primer annealing can occur due to these low temperatures. This nonspecific annealed primer can then be extended by the Taq DNA polymerase, generating nonspecific products and lowering product yields. Hot Start PCR significantly reduces nonspecific priming, the formation of primer dimers, and often, increases product yields[citation needed]. In Hot Start Long and Accurate PCR, the impact on yield can be dramatic[citation needed]. Classic methods, while effective, involve additional handling and increased risk of contamination. [1] Source of the article published in description is Wikipedia. I am sharing their material. Copyright by original content developers of Wikipedia.
Link- http://en.wikipedia.org/wiki/Main_Page
11 سال پیش
در تاریخ 1392/08/02 منتشر شده
است.
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