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Question about Sequencing

Genetics as it applies to evolution, molecular biology, and medical aspects.

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Question about Sequencing

Postby xiaolong88 » Thu Oct 14, 2010 4:39 pm

Honestly there are a lot of DNA sequencing technique, but what is the method used to sequence or to detect a shorter DNA fragment which is extremely small probably <100bp. As i know Sanger sequencing can read up to >500bp.
Would it be illumina sequencing?
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Postby lishenghui » Thu Oct 14, 2010 5:36 pm

It is widespread for sequencing of short fragment in illumina machines within the reads length of ~100, ~75 or ever ~40.....
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Postby xiaolong88 » Thu Oct 14, 2010 5:57 pm

Thanks for your info. Yes i did read an article about it but yet i can't find any book which explain about this method. Anyway thanks for your reply
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Postby canalon » Thu Oct 14, 2010 8:40 pm

The new methods (SOLiD, Helicos, 454 Illumina etc) allow massive numbers of simultaneous short reads (30/40 up to 100 for illumina but with less reads per reaction).
Sanger allow much longer reads (700 up to 1000 in a good day), but of one fragment at a time. In terms of cost the next gen sequencing is not interesting to sequence one gene/fragment at a time, however short because of the massive build-in redundancy.
So it depends what you want to do.
An introduction of the methods can be found on the different manufacturer websites or briefly here:http://en.wikipedia.org/wiki/Sequencing (see pyrosequencing, but helicos is not presented, you will have to look for them)
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Postby xiaolong88 » Fri Oct 15, 2010 5:46 am

Thanks Canalon

Yes i did go through the article online include the wiki and books(most trusted). I know that there are a lot of methods used in sequencing. But what about IP-RP-HPLC, ion-pair reversed-phase high-performance liquid chromatography. Does this method can used to read short fragment of DNA sequences as well?
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