Talk:Pyrosequencing

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Untitled[edit]

The subsequent word's are my own and nobody else's. I re-wrote them after supposed copyright infringement. The encyclopedia will be missing a huge area of development without them. Use them or don't use them, either way i'm not bothered.

Pyrosequencing, a new method of DNA sequencing which involves the addition of phosphorylated dNTP's to the DNA polymerase reaction which emmit light upon binding to the DNA template. It makes DNA sequencing significantly faster than with the current chain termination method.

We do use them, look at Pyrosequencing/Temp. The text there will replace the text currently at Pyrosequencing as soon as that article is deleted completely. Sorry for that inconvenience, this is necessary because otherwise, the copyrighted text will remain visible via the articles history. I already wrote a welcome message to you at your previous ip's talk page, but obviously you didn't see it. So again: Welcome here, it's nice to have a knowledged molecular biologist here :)))) . Please do not feel put off wikipedia, you'll soon get used to our customs if you read some of the guidelines and keep editing. If you have questions, leave a message at my talk page.

I strongly suggest you get a user name. It's free, easy and anonymous: Then it'll be easier to communicate with you politely and efficiently.

READ LENGTH: Is it not so that 454 with the FLX+ chemistry is capable of read lengths of up to 850 nucleotides matching that of Sanger sequencing?

Again:welcome!

Lady Tenar 13:30, 13 Nov 2004 (UTC)

Did you mean "emmit" light or "emit" light? — RJH 19:50, 9 Feb 2005 (UTC)

Zaimon 12:47, 3 May 2007 (UTC)

I edited this article to try and make it more comprehensible for the average reader. Is it alright?

It's definitely all right. As Wikipedia says, Be Bold! This page has needed work for a while. I'll get around to helping out one of these days. Spin2cool 18:32, 4 May 2007 (UTC)

After reading the introduction I feel that the 454 should only be mentioned later under recent advantages. This way it appears that 454 is the only way pyrosequencing is applied. In addition, it is a bit of a repetition.CharonZ 10:06, 10 July 2007 (UTC)

To improve[edit]

I wasn't be able to understand with this article, how you mesure which lettre you detected, that you do it in different containers. Maybe you could edit it in any way? Here is a page where it is discribed well as well, maybe you can add it :-) http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/P/Pyrosequencing.html —Preceding unsigned comment added by 134.59.6.84 (talk) 16:45, 18 January 2011 (UTC)

I'm not exactly an expert, but this article seems fine to me[edit]

Remove the 'article needs an expert' tag? Its a garbage tag anyway if you ask me. I am a graduate in Biochemistry, and the article reads fine to me. (I didn't go into the details in detail). --Dan|(talk) 14:59, 4 March 2008 (UTC)

Agreed. The template has been there for a year and the article seems fine to me as a regular user of the method, I'm removing it. If anyone has any ideas for improvement (other than the intro which I might rewrite myself), please tell us. AstarothCY (talk) 16:28, 4 March 2008 (UTC)
'can generate 400 million nucleotide data in a 10 hour run with a single machine. Each run would cost about 5,000-7,000 USD, pushing de novo sequencing of mammalian genomes into the million dollar range.'

surely this is wrong, if human dna has 6bill base pairs then you need in the region of 15 runs. placing the cost of sequencing the in the region of 75000 - 100000 usd not near the million dollar range as stated. however if multiple runs are required, say a 7-8 fold coverage, then the cost would reach the stated region of 1mill usd. the statement should be changed accordingly —Preceding unsigned comment added by 86.164.137.107 (talk) 00:40, 22 November 2010 (UTC)

Too obscure??[edit]

The article is clear, I believe it can be easily understood by an undergrad in life science and pyrosequencing is not high school stuff. I personally would remove the parenthesis after DNA sequencing which is plain silly: evryone knows what DNA sequencing is as they sequenced the human genome! --Squidonius (talk) 17:29, 15 April 2008 (UTC)

Suggestion[edit]

It took me a while to understand why the dNTPs - once they are added - do not bind to every other corresponding base in the whole strand. I'm assuming now that this is not the case, because the polymerase needs a 3'-binding site to build in a nucleotide. If my assumption is correct I suggest that somebody with a more precise scientific vocabulary should add a passage to clearify this part. Cheers and good night LeuschteLampe (talk) 20:16, 11 April 2013 (UTC)

How Pyrosequencing Works (Graph added)[edit]

I've just added a SVG graph showing the whole procedure. Feedbacks appreciated, thanks! Thisisjp (talk) 16:38, 1 December 2014 (UTC)

Just been puzzling through it, and I think I can follow it all, with one exception. It seems that the apyrase, which mops up unused dNTPs does its job by breaking the dNTP down to dNMP + PPi. Surely then the PPi would kick off the same luciferase-catalysed reaction that indicate incorporation? The only way I can see of avoiding this is segregating the apyrase and its breakdown products from the sulfurylase/luciferase, but this doesn't seem to be mentioned. 82.173.146.134 (talk) 19:40, 4 August 2018 (UTC)

History and relevance of pyrosequencing[edit]

There is, at the least from my side, a bit of confusion. Is pyrosequencing still in use? Some text books claim so, but elsewhere pyrosequencing has been replaced in NGS more or less completely. On the other hand I have just read that variants of pyrosequencing may still be in use, albeit in different ways or improved variants? No clue. Either way, I think it would be useful to add a history subsection to the article, because while I think pyrosequencing is no longer any massively used sequencing strategy, it is possibly not completely dead, more like semi-buried or has changed use patterns, even with the general decline. 2A02:8388:1604:CA80:5EA1:25A:77B5:389E (talk) 17:05, 26 June 2019 (UTC)