Talk:Real-time polymerase chain reaction

message: Dear PCR gurus, I try to find the answer by reading this wiki page.
"It monitors the amplification of a targeted DNA molecule during the PCR (i.e., in real time), not at its end, as in conventional PCR."
Why scientisits need such information? Does it mean that knowing how fast the RNA being copied relate some facts about the sample?  — Preceding unsigned comment added by Randomwangran (talk • contribs) 17:14, 24 January 2020 (UTC)
thread_title:  why scientisits need real-time data? 

message: Does real-time PCR use FRET or some other kind of energy transfer? Heid's paper "Real Time Quantitative PCR" only refers to it as "fluorescent energy transfer." However, the wiki page on FRET states that it is NOT in fact fluorescence as it is not a "radiative" energy emission. Any help clearing this up?    —Preceding unsigned comment added by 146.189.246.13 (talk) 15:20, 16 May 2008 (UTC)
replies: text: Sure. TaqMan is based on FRET. here is a free example. There are many tens of such examples. Tzachi Bar (talk) 10:10, 12 November 2009 (UTC)
thread_title: Confusion on methods

message: While the images look very pretty, they are also pretty inaccurate. The DNAs depicted are left-handed and have no major or minor grooves. I don't know if aesthetics or complete accuracy is more important here. I've also moved one image, since it had been badly integrated into the text. --163.1.176.254 14:25, 25 May 2007 (UTC)
thread_title:  Images 

message: Needs merging with Real-time PCR/Temp —Preceding unsigned comment added by Zephyris (talk • contribs) 
:Merging done. Elle vécut heureuse à jamais (Be eudaimonic!) 23:47, 16 February 2006 (UTC)
thread_title: Merge with real-time PCR/Temp

message: Much of the information on this page seems more related to reverse transcription pcr - I understand real time pcr has applications in fields such as ancient DNA and forensics? Aaadddaaammm 22:27, 31 January 2007 (UTC)
replies: text: There seems to be a confusion of acronyms: RT-PCR is either "Real Time" PCR (Quantitative) or "Reverse Transcriptase" PCR (Qualitative PCR for RNA), I think we need to distinguish these concepts.
Oderbolz 08:15, 21 February 2007 (UTC)
thread_title:  Reverse transcription pcr? 

message: I just added a flash video on various methods of fluorescence and quenching to the external links section. It might be wikispam, but to me at least, it seems more informative than the other animation in the links right now. Ryan Brady (talk) 19:03, 3 November 2009 (UTC)
thread_title:  Real-time PCR animations 

message: Just wondering why the pointer to MIQE guidelines was removed?  If you're doing any academic work involving rtpcr they're very relevant.
SamMason79 (talk) 18:17, 17 February 2011 (UTC)
replies: text: Hello SamMason79, in general, any information from the literature that is relevant to the entry should be included in the article, not referred to by external links. Especially entries like this one seem to attract an inordinate number of external links over time, to the point that the list becomes very unwieldy. Therefore, when it comes to ELs, less is more. Please consult WP:EL for what external links are acceptable, and if you still think this link should be added please discuss here. Thanks. Malljaja (talk) 23:25, 17 February 2011 (UTC)
thread_title:  MIQE guidelines 

message: The article sas:
"breakdown of the probe by the 5' to 3' exonuclease activity of the Taq polymerase breaks the reporter-quencher proximity and thus allows unquenched emission of fluorescence"
but when you see the Taq polymerase article it says:
"One of Taq's drawbacks is its relatively low replication fidelity. It lacks a 3' to 5' exonuclease proofreading activity"
If the Taq polymerase doesn't have 3' to 5' exonuclease activity, then how is the reporter-quencher proximity broken?
Ammaralsheik (talk) 10:37, 7 July 2011 (UTC)
thread_title:  Apparent mistake under "Flourescence reporter probe method". 

message: I arrived here by trying to find out if mRNA expression in cells could be monitored in real time (and if so at what 'frame' rate).
After reading the article it seems like it is really just a more accurate and linear measurement of PCR amplification (when nor measuring at intermediate stages in PCR process some species will saturate at a plateau level and hence we have nonlinear distortion in quantity/concentration). This explains the naming qPCR, however it does not explain the naming Real-time (it IS in a way "real time" with respect to the PCR process as a technique but not to the use of the technique, i.e. monitoring mRNA expression levels of a cell in real-time).
Below I read that 2 articles were merged: qPCR and real-time PCR.
It seems like either the real-time naming variant is either biotech marketing hype (in which case the 2 articles were correctly merged and the article should express caution around the misleading misnomer) or there really is such a thing as real-time PCR, as a kind of qPCR but which can be performed realtime to monitor a cells mRNA expression levels. Calling something real-time necessitates a discussion about current technological data rates. i.e. email can be considered real-time compared to snail mail. But instant messaging can be considered more realtime than email...
I dont see how making measurements during PCR stages make it happen faster, only the quantity more precise by having measured the last nonsaturated quantity in a specific cycle of the PCR reaction before this species is saturated... In short the article explains how to increase concentration resolution, but not time resolution.— Preceding unsigned comment added by 83.134.175.158 (talk) 16:15, 2 December 2011 (UTC)
replies: text: The term "real-time PCR" and its variant "qPCR" are very widely and interchangeably used in molecular biology research. "Real time" refers to the acquisition of data during the PCR process, i.e. at each cycle (rather than a single end-point measurement). Measuring accumulation of mRNA in living cells in "real time" is possible with this technique as well, for example, by monitoring expression in a time course (i.e., sequential removal of cells over time for reverse-transcription real-time PCR analysis). You may note that Wikipedia follows the usage convention established in the literature, even if some of the terminology may be confusing. Malljaja (talk) 16:58, 2 December 2011 (UTC)
replies: text: Well what you describe is still not real time gene expression measurement: you are measuring mRNA expression levels of different cells of which you assume that if they are as old will pass through the exact same states...  — Preceding unsigned comment added by 134.184.49.148 (talk) 12:00, 6 December 2011 (UTC)
replies: text: I'm not quite sure what you're trying to say here, but I take it that you are talking about synchrony of mRNA expression. This is a physiological phenomenon which may be solved by synchronizing populations of cells, so that they are all in the same cell cycle. This is a biological challenge, whereas real-time PCR is a technical method to measure accumulation of a target nucleic acid (be it a piece of cDNA from mRNA or genomic DNA) in real time. The method is not predicated on synchronized expression of genes of interest—it looks at the presence of one or more transcripts across populations of cells or even within a single cell, depending on how the experiment is set up. Malljaja (talk) 15:06, 13 December 2011 (UTC)

text: I too have come here looking for a "duration": how long does the technique take to output the information? from the perspective of a complete outsider, I was led to believe by the name "real-time" that technology got so fast as to be able to output some form of genetic information in "real-time", or as soon as / shortly after the sample is deposited in a machine. Probably a clarification on the name, and duration of the process would make the information more accessible to the general public, or at least less ambiguous. thanks  — Preceding unsigned comment added by 79.159.111.244 (talk) 09:32, 30 January 2020 (UTC)
thread_title:  "Real-time" PCR