Friday, June 20, 2008

454

I have been trying to understand the pyrosequencing technology, popularly known as 454. The very first question which came to my mind is WHY it is called 454? Is it because .454 kasool Handgun is sleeker then Shotgun ( Cf. Whole Genome Shotgun Sequence ) and slowly becoming as powerful ... or is it because of the fact that "In one instrument run sequence a minimum of 20 million base pairs in 4.5 hours" ... just a conjecture, if you guys have a proof, please share !
Anyways coming back to the technology, what I understand (below is a crude attempt to visualize it) is that there are 4 containers containing A,T,G and C which sequentially pass over the bowl containing a mixture of Polymerase, Sulfurylase, Luciferase, Apyrase and the DNA fragment to be sequenced. The addition of the nucleotide will light up ( thanks to lucifer-ase ) the CCD (a thing close to what we have in a digital camera and the costliest part of the instrument). Since we know the culprit nucleotide container, peak telling how many of that nucleotide is added, we can sequentially proceed and keep adding letters to the reads... this is happening in a massively parallel fashion and that brings out the power of 454...



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