Talk:Real-time polymerase chain reaction

why scientisits need real-time data?

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)

Confusion on methods

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)

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)

Images

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)

Merge with qPCR?

A lot of duplication exists between qPCR and this page. Admittedly they are not exactly the same, but some links should be made between the two. Any comments? Dr Aaron 23:05, 1 August 2006 (UTC)

I am not an expert, but I think that the "Fluorescent reporter probe method" of qPCR is the same as RT-PCR using a probe with reporter and quencher.

Oderbolz 08:12, 21 February 2007 (UTC)

I just finsihed redirecting qPCR to this page. Should someone add a comment on the page about this? --Michael (talk) 17:36, 11 June 2008 (UTC)

After the merge, the name really should be "qPCR," since nobody calls it RealTime-PCR in journals, they call it RT-qPCR, and nobody ever refers to it as "RT-PCR" because of the confusion with Reverse Transcriptase coupled PCR. All qPCR is in Real Time. The MIQE guidelines note on the article itself really bears this out: "The MIQE guidelines propose that the abbreviation qPCR be used for quantitative real-time PCR and that RT-qPCR be used for reverse transcription–qPCR"--Shibbolethink (talk) 16:02, 7 January 2015 (UTC)

Merge with real-time PCR/Temp

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)

Reverse transcription pcr?

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)

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)

Real-time PCR animations

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)

MIQE guidelines

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)

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)

Apparent mistake under "Flourescence reporter probe method".

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) The 5'->3' exonuclease activity degrades DNA downstream, that is already hibridized to the template as the Taq advances. It is used in the nick translation technique. The 3'->5' activity degrades the last polymerized nucleotide if it is a mismatch, and serves for proofreading. — Preceding unsigned comment added by 190.247.87.220 (talk) 14:46, 8 November 2012 (UTC)

"Real-time" PCR

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)

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)

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)

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)

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)

Requested move

The following discussion is an archived discussion of the proposal. Please do not modify it. Subsequent comments should be made in a new section on the talk page. No further edits should be made to this section.

The result of the proposal was no consensus. --BDD (talk) 16:56, 27 August 2013 (UTC)

Real-time polymerase chain reaction → Quantitative polymerase chain reaction – Both names are used but this one is more precise and by far easier to understand. What is more, the abbreviation used to name this technique is qPCR (to avoid confusion with Reverse transcription polymerase chain reaction, see article). Relisted. BDD (talk) 18:25, 20 August 2013 (UTC) Stormpananal (talk) 16:39, 11 August 2013 (UTC)

• Support strongly for these reasons. Also, many articles used the unnecessarily long name "quantitative real-time PCR". But "quantitative PCR" is the common name ; and it carries all the necessary information to know which is the technique and what it does. Hatagalow (talk) 16:49, 11 August 2013 (UTC). Sockpuppet of proposer.
• Support for the same reasons. Daleslimnsaw (talk) 19:14, 11 August 2013 (UTC).Sockpuppet of proposer
• Support because it is the logical and common name. Also it matches the qPCR recommended abbreviation. Rumbartangala (talk) 19:54, 11 August 2013 (UTC). Sockpuppet of proposer.
• Oppose They are similar techniques but they are not the same. Also see Wikipedia:Sockpuppet investigations/Rumbartangala. Graham Colm (talk) 20:14, 11 August 2013 (UTC)

A real-time PCR is a quantitative PCR. As the article is on quantitative PCR in general, the title should be quantitative PCR. Triolysat (talk) 13:53, 15 August 2013 (UTC).

The article is not about quantitative PCR in general. It only describes the type of quantitative PCR known as "real-time PCR." It would be much easier to keep the current article title here and have a new article describing the different forms of quantitative PCR. CatPath (talk) 19:21, 15 August 2013 (UTC)

• Oppose strongly. Real-time PCR is merely one approach to qPCR. Other methods of qPCR were performed before real-time PCR became the favored method. CatPath (talk) 02:23, 13 August 2013 (UTC)

What you say is true but speaks if favour of the new title! Triolysat (talk) 13:51, 15 August 2013 (UTC).

I disagree with Triolysat . Graham Colm (talk) 21:56, 15 August 2013 (UTC)

• Support strongly. Real-time PCR is merely one approach to qPCR. The article is on quantitative PCR in general. The links that redirect here are mostly "quantitative PCR" and for all these reasons it should be the title. Triolysat (talk) 13:51, 15 August 2013 (UTC).

The article in its current form is not about quantitative PCR in general. There is no mention of other methods of quantitative PCR. Only variants of real-time pcr are described. Therefore, this article should retain its title, and there should be a separate "quantitative PCR" article. CatPath (talk) 16:51, 15 August 2013 (UTC)

I agree with CatPath. Graham Colm (talk) 21:56, 15 August 2013 (UTC)

You could both create this article (quantitative PCR) together if you think this is the best option. If not, renaming would be the thing to do. Because most people that arrived on this article did so by clicking on a quantitative PCR link. And before creating a new article, see the sections above: real-time PCR and qPCR were merged together few years ago for good reasons. I am not sure doing the opposite is useful for readers. Triolysat (talk) 07:00, 16 August 2013 (UTC).

• Support strongly. As stated above on this page, "The term 'real-time PC' and its variant 'qPCR' are very widely and interchangeably used in molecular biology research". Many people consider these two terms as synonymous. And real-time PCR is abbreviated qPCR so any distinction tend to disappear. The article should use the most common, most logical and most used term. Seamsurgegen (talk) 07:10, 17 August 2013 (UTC).

I don't agree that real-time pcr and qPCR are considered synonymous, at least in formal usage. Take a look at the titles of the articles listed in the References section. When pcr is mentioned, most use the wording "real-time quantitative pcr" or some similar wording that retains the term "real-time." The exception is the Biotechniques article, the first paper cited (in the current version of the article). Even there, they use the words "real-time qPCR" when the technique is first referred to in the abstract. The title "quantitative pcr" is misleading since there are other forms of quantitative pcr. CatPath (talk) 20:32, 19 August 2013 (UTC)

• Relisting comment Given the sockpuppetry that's already occurred in this request, it would be helpful to get more outside voices to help build consensus. --BDD (talk) 18:25, 20 August 2013 (UTC)

The above discussion is preserved as an archive of the proposal. Please do not modify it. Subsequent comments should be made in a new section on this talk page. No further edits should be made to this section.

Requested move 7 January 2015

The following is a closed discussion of a requested move. Please do not modify it. Subsequent comments should be made in a new section on the talk page. Editors desiring to contest the closing decision should consider a move review. No further edits should be made to this section.

The result of the move request was: not moved. No consensus on whether the move is a good idea – despite being left open for over a month, only two editors have commented, and I don't really see a winner in terms of the arguments put forward. Number 57 14:35, 12 February 2015 (UTC)

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Real-time polymerase chain reaction → Quantitative PCR – This discussion has happened once before, but I really think it's worth having again. Colloquially, in the field, no one refers to it as Real-Time PCR. See here and here. Plus, under Wikipedia's article title conventions, the name should describe current usage. Also in those guidelines is a small bit about using titles to help distinguish similar concepts. RT-PCR in the field means Reverse transcriptase-PCR. Real-Time PCR is NEVER used in journals or in conversation as a result. It's all qPCR. It's all Quantitative PCR. Plus this article is entirely ABOUT qPCR. New students learning about these concepts are only confused by the current title. --Relisted.  — Amakuru (talk) 21:27, 15 January 2015 (UTC) Shibbolethink (talk) 16:15, 7 January 2015 (UTC)

Never? Look at the References. I see several that use "real-time PCR" or "real-time RT-PCR" in the title. Also, the very first link you provide uses "real-time PCR" in its title. CatPath (talk) 17:59, 7 January 2015 (UTC)

My main objection to renaming the article "Quantitative PCR" is that this article is solely about the real-time method for quantifying nucleic acids. There are other PCR-based methods for quantifying nucleic acids, including competitive PCR and digital PCR. Changing the title to "Quantitative PCR" would mean that we would have to include descriptions of these other methods of quantitative PCR in this article. CatPath (talk) 18:29, 7 January 2015 (UTC)

Those references all use "qPCR" as the abbreviation. What does "qPCR" stand for? I fail to see how expanding this article to include all methods of qPCR is a bad thing. I think, if this article is going to use the term "qPCR" as synonymous with "real-time PCR," then it should encapsulate all of "Quantitative PCR." Either that or it should be split into two articles, one entitled "Real-Time PCR" and one entitled "Quantitative PCR" and over time, the first article will fall into disuse, while the second is pretty much this article plus all other methods of qPCR. Why keep that information out of Wikipedia? Why not describe what qPCR is at large?--Shibbolethink (talk) 18:46, 7 January 2015 (UTC)

Here are several examples of papers in the highest impact journals using "Quantitative PCR" as the colloquial expression in their results and discussions. These are all from recent years. 1 2 3 4 5 6 7--Shibbolethink (talk) 19:07, 7 January 2015 (UTC)

I agree that "qPCR" is used colloquially by some to refer to real-time PCR (although I hear "real-time PCR" >90% of the time where I'm located). According to one of the links you provided, Wikipedia guidelines favor "common names," but not when the common name lacks the precision to describe the scope of the article. And that's the problem with the title "Quantitative PCR." In my opinion, the current title should remain. It doesn't sound like a good idea to have separate articles for the reasons you state. I also disagree with having a single all-encompassing "Quantitative PCR" article since the real-time method deserves its own article. To accomodate other approaches to quantitative PCR, a "History" section can be added where older techniques such as competitive PCR would be described. There's already an article about digital PCR. CatPath (talk) 22:56, 8 January 2015 (UTC)

• Support move per the sources above and to reduce confusion with reverse transcription PCR. (FWIW, I hear "qPCR" >90% of the time where I'm located!) No prejudice against making a new article at this title as long as it makes the distinction clear, but if there will be only one article then I think it should be at qPCR. Sunrise (talk) 22:10, 23 January 2015 (UTC)

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The above discussion is preserved as an archive of a requested move. Please do not modify it. Subsequent comments should be made in a new section on this talk page or in a move review. No further edits should be made to this section.