Illumina (Solexa) sequencing

Further Illumina dyes sequences is a technique used to determine the sequence of base pairs of DNA, known as DNA sequencing. Solexa and Manteia prediction medicine, it has been developed by scientists from the acquired companies to Illumina later. This sequence is based on the method of thermally reversible terminator that allows the identification of single base thereof, as introduced into the DNA strand. Repeat sequences homopolymers and often, it is used for local sequence difficult. Further, the protein field transcriptome and genome-wide, small RNA discovery methylation profiling, whole genome – it can be used for sequencing of the nucleic acid interaction analysis.

Part of the now Solexa, Illumina, a method for technology reversible dye terminator sequencing-based, it has developed the engineering of polymerases have developed internally. Solexa Solexa system design was invented by Klennerman and Balasubramanian of the Chemistry Department at the University of Cambridge in was developed internally [49] wound chemistry. The company has acquired the Solexa Manteia prediction drugs that massivelly get the parallel sequencing technology based on “DNA cluster” containing the cloned DNA amplification on the surface in 2004. Cluster technology was acquired in the treatment of joint California Lynx. Solexa, Inc. merged with Solexa’s form of Lynx later

In this method, primers and DNA molecules, are applied to the first slide, DNA of the local clone colonies were amplified by polymerase minted later “DNA cluster” to be formed. The sequenced, four kinds of bases reversible terminator (RT-based) was added, and nucleotides unincorporated is eluted. Camera is removed from the DNA that makes it possible to get the image of nucleotides that are fluorescently labeled, and starts the next cycle chemically dyes with terminal 3 ‘blocker thereafter. Unlike pyrosequencing, DNA strand extending one nucleotide and image acquisition time may be performed in time slow allows large arrays very DNA colonies should be covered by Continuous shooting from a camera.

Separation and shooting enzyme reaction allows unlimited capacity Thing in theory and optimal performance. The optimum configuration of an analog in this manner eventually – determining the amount of the chamber multiplied by the number of cameras in only digital conversion pixel of DNA colonies required (approximately 10 pixels for optimum viewing thereof colonies reach divided by the number, the performance of the instrument). In 2012, after working in enzymatics optics, and fluid camera is available with A / D conversion speed MHz, more than 10, is a multiple of one million nucleotides  corresponds to 1X to about one human genome equivalent bandwidth coating of the human genome sequence at the time and again on one instrument (camera) equipment the day it is possible.

The basic steps for sequencing Illumina dye response is as follows. DNA molecules is attached to the primer first slide, I was amplified colonies local to be formed. This technology 1998 Laurent Farine Li and Geneva Medical Glaxo Wellcome Institute Dr. Pascal Mayer Dr. cluster generation was invented, has been developed, was presented to the public for the first time in the O end of 1996. Labeled with a fluorescent color different termination bases (adenine, cytosine, guanine, and thymine) reversibly four types, is added associated with the blocking group. Then, the four bases were then compete for binding sites of the template DNA for removing molecules sequenced, are not captured. Each laser after fusion is applied as a result of the probe and the removal of the terminal 3 ‘blocking groups. Then, to enable the start of the next cycle and identification sequence, the identification of detectable fluorescent color for one of the four bases were observed. Until sequencing the complete DNA molecule, the process is repeated.

This technique provides a number of advantages over traditional methods of sequencing by Sanger sequencing. I get a real data series quickly and can be due to the nature of the sequence Illumina dye that is automated, to thread multiple sequences at the same time. In addition, in contrast to the enzyme very expensive and required by sequencing other, this method uses a DNA polymerase only.