Real time PCR using taqman dye

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A quantitative polymerase chain reaction (qPCR), also called real-time polymerase chain reaction, is a laboratory technique of molecular biology based on the polymerase chain reaction (PCR), which is used to amplify and simultaneously quantify a targeted DNA molecule. For one or more specific sequences in a DNA sample, quantitative PCR enables both detection and quantification. The quantity can be either an absolute number of copies or a relative amount when normalized to DNA input or additional normalizing genes.

The procedure follows the general principle of polymerase chain reaction; its key feature is that the amplified DNA is detected as the reaction progresses in "real time". This is a new approach compared to standard PCR, where the product of the reaction is detected at its end. Two common methods for the detection of products in quantitative PCR are: (1) non-specific fluorescent dyes that intercalate with any double-stranded DNA, and (2) sequence-specific DNA probes consisting of oligonucleotides that are labelled with a fluorescent reporter which permits detection only after hybridization of the probe with its complementary sequence to quantify messenger RNA (mRNA) and non-coding RNA in cells or tissues.

qPCR is the abbreviation used for quantitative PCR (real-time PCR).[1] Real-time reverse-transcription PCR is often denoted as: qRT-PCR[2][3][4] The acronym "RT-PCR" commonly denotes reverse transcription polymerase chain reaction and not real-time PCR, but not all authors adhere to this convention.[5] Source of the article published in description is Wikipedia. I am sharing their material. Copyright by original content developers of Wikipedia.

[capriceporter]

You really helped me out on this one, I thought it would have been a bit complicated when I looked at lecture slides. THANKS!

[simonwu]

I was firstly surprised that Shomu's Bio got "this" deep into such an "unpopular" topic, while I then realized I was so naive to be called a bio student because I am realistically way far to professional.

[ithirstyforknowledge]

question: since there will be many more quenchers in the solution, why will other quencher not inhibit the reporter?

[sanjeetk]

Boss please dont created confusion
1.Quencher: Dye release photon after the excitation due to applied energy ( Light) and Quencher not inhibit the dye to emit the photon. Its is phenomenon called FRET( Energy transfer from dye to quencher if its very close to each other.
and the dye choped by polymerase by exonuclease activity...Thanks
 

[shoxankader]

i want to know is taqman mean probe based qPCR  or they are differe 

[shaheentarannum]

What is the principle of real time pcr taqman

[thurayae]

please shomu, do a video for taqman SNP allelic discrimination, analysis for data

[pawarjay]

cleavage(5' exonuclease)based/Taqman

[pramitbanerjee]

you need to put these videos in the proper order..