Real-Time PCR

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تمامی اصول و واکنش‌گرهایی که برای یک RT-PCR معمولی نیاز است دراین تکنیک هم بکار می‌رود اما یک گزارشگر فلورسنت نیز در واکنش حضور دارد. این گزارش‌گرها به گونه‌ای طراحی می‌شوند که در صورت تکثیر DNA نور تولید کنند. لذا افزایش شدت نور ثبت شده در دستگاه با میزان محصول بدست آمده نسبت مستقیم دارد.با ادامه یافتن PCR شدت فلورسنت رو به افزایش می‌گذارد.
به اولین چرخه‌ای که شدت فلورسنت بیشتر از خط پایه باشد چرخۀ آستانه یا CT گویند. عدد CT با مقدار الگوی اولیه رابطۀ معنی‌دار دارد و از روی آن می‌توان مقدار mRNA اولیه را تخمین زد. به عبارت دیگر در فاز اولیه مرحله تصاعدی مقدار فلورسنت افزایش یافته و به آستانه ­ای می رسد که به مقدارمشخصی از سطح background بالاتر است، این چرخه ( سیکلی از PCR که قطعه تکثیر از حد آستانه عبور می کند) به عنوان CT شناخته می شود که در برخی منابع با عنوان Crossingpoint مطرح شده است. این مرحله شروع نسخه برداری از قالب است که در محاسبات نتایج آزمایش استفاده می شود...

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1394/06/02

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لینک منبع :Real-Time PCR

A real-time polymerase chain reaction is a laboratory technique of molecular biology based on the polymerase chain reaction (PCR). It monitors the amplification of a targeted DNA molecule during the PCR, i.e. in real-time, and not at its end, as in conventional PCR. Real-time polymerase chain reaction - Wikipedia, the free ... https://en.wikipedia.org/wiki/Real-time_polymerase_chain_reaction Real-time polymerase chain reaction - Wikipedia, the free ... https://en.wikipedia.org/wiki/Real-time_polymerase_chain_reaction‎Cached SimilarA real-time polymerase chain reaction is a laboratory technique of molecular biology based on the polymerase chain reaction (PCR). It monitors the amplification of a targeted DNA molecule during the PCR, i.e. in real-time, and not at its end, as in conventional PCR. ‎Background - ‎Basic principles - ‎Classification - ‎Fusion temperature analysis[PDF] Beginner's Guide to Real-Time PCR - Primerdesign Ltd www.primerdesign.co.uk/.../beginners_guide_to_real_time_pcr.pdf‎Cached SimilarTo understand real time PCR it is easier to begin with the principles of a basic PCR: PCR is a technique for amplifying DNA. There are 2 reasons why you may ... Images for Real-Time PCR What is Real-Time PCR (qPCR)? | Applications & Technologies | Bio ... www.bio-rad.com/en-us/applications.../what-real-time-pcr-qpcr‎CachedAn array of solutions to optimize, carry out, and troubleshoot MIQE-compliant real -time PCR experiments. Introduction to Real-Time PCR Primer Design - PREMIER Biosoft www.premierbiosoft.com/tech_notes/real_time_PCR.html‎Cached SimilarAt the same time, a relatively small amount of PCR product (DNA, cDNA or RNA) can be quantified. Real Time PCR is based on the detection of the fluorescence produced by a reporter molecule which increases, as the reaction proceeds. This occurs due to the accumulation of the PCR product with each cycle of amplification. [PDF] Real-Time PCR Vs. Traditional PCR - Applied Biosystems www6.appliedbiosystems.com/support/tutorials/.../rtpcr_vs_tradpcr.pdf‎Cached SimilarDNA Extraction. PCR. Real Time. Detection. Real-Time Vs Traditional PCR. Real -Time chemistries allow for the detection of PCR amplification during the early ... [PDF] Real-time PCR handbook - Gene-Quantification.info www.gene-quantification.com/real-time-pcr-handbook-life-technologies-update-flr.pdf‎Cached SimilarIn real-time PCR, the amount of DNA is measured after each cycle via ... fluorescence against the cycle number, the real-time PCR instrument generates an ... [PDF] Real-Time PCR Applications Guide - Gene-Quantification.info www.gene-quantification.com/real-time-pcr-guide-bio-rad.pdf‎Cached Similarof real-time PCR in specific applications, namely, gene expression analysis, allelic discrimination ... Real-time PCR that is quantitative is also known as qPCR . qPCR Basics - Biosistemika biosistemika.com/workshops/qpcr-basics/‎Cached SimilarThe method of choice for nucleic acid (DNA, RNA) quantification in all areas of molecular biology is real-time PCR or quantitative PCR (qPCR). The method is ... Real-Time PCR (qPCR) | Thermo Fisher Scientific https://www.thermofisher.com/us/en/home/.../pcr/real-time-pcr.html‎Cached SimilarReal-time PCR is a form of polymerase chain reaction (PCR) where data are collected in real-time as the reaction proceeds. Continuous data collection enables ...