Advanced methods

In many cases, sequencing large the entire sequence, part a very long and full chromosomal DNA, a large array is used to generate short sequence, for example, as seen in phage display, a lot of very may be. For many purposes, such as chromosome, such as this, the general approach, (with mechanical forces) and shear (by restriction enzyme) cutting, consists of a large fragment of DNA into short fragments of DNA are. It can be cloned into a vector DNA is then fragmented into DNA, is amplified in a bacterial host such as E. coli. Are arranged in a sequence of these individual, DNA fragment purification short, assembled electronically into one long from bacterial colonies each.

The term “de novo sequence” refers to a method used to determine the sequence DNA sequence is not known initially. Gaps in the sequence assembled de novo conversion from Latin to “. Beginning” may be filled by primer walking. The different strategies, has a different trade-off speed and accuracy, the method of the gun is used to sequence the genome often large, and assembly, causing the failure of the assembled genome frequently arranged Te is repeated, in particular, is complicated and difficult.

Advanced methods

As a method of detecting these molecules, the sequence of approaching the most used in the in vitro cloning steps to amplify the DNA molecules of the individual, not sensitive enough for the molecular sequencing single. The emulsion PCR, isolated DNA molecules of beads coated with a primer in the liquid droplets in the oil phase. Polymerase chain reaction (PCR), each bead of copies were cloned DNA molecules were fixed for preservation sequencing the court. Emulsion PCR is used in a way that has been developed to Marguilis etc.. Etc. Porreca, and Shendure (commercialized by 454 Life Sciences). (Also known as “Bologna sequence”), solid sequencing.

Have professional installation of sequencing and genome sequencing, a novel to read for the first time will involve sequencing new de. Unique combination of the length of the reading, to read the size of the flexible insert paired end and depth, making it ideal for Illumina sequencer for de novo sequence determination. Accuracy of reading raw unprecedented provides efficient production reliability and contigs, high-quality long assembly. In review, method for the de novo sequencing of peptides by mass spectrometry is described. De novo method uses the approach of computations to derive the sequence or a partial sequence of the peptide directly from the MS / MS spectrum experimental. Concept behind the new sequencing method in December has been described. Another method for identifying peptides by tandem mass spectrometry corresponding to the fragment ions of the virtual peptide ions produced by genomic or protein database. How to de novo is essential for the identification of proteins genome is not known, but they will not be affected by an error in the search database, even when the genome is known, they are It is also particularly useful. Another advantage of the de novo method may be used to search for mutations or post-translational modifications that identified by the software for sequence homology based partial.

New generation of sequencing technology has revolutionized molecular biology. Applied Biosystems’ solid generated gigabases nucleotide sequence week and Illumina Solexa. It will nevertheless be perceived limitations of ultra-high-throughput techniques these is the length read their short. Research is a field of mature Unfortunately, the sequence assembly of existing programs, that of reads generated by classical Sanger capillary sequencing of de reassembly sequence is valid for short sequence that is generated by reading Illumina In said the platform that it was not. In early studies, many gaps have suggested to provide can be used for generally 20 to 30 nucleotides to useful products, a short sequence that can be said that includes prokaryotes and eukaryotes assembly of the genome sequence of the. To read from 30 to 50 nucleotides in length, a software tool that can be used freely with the objective assembly clearly evidence and principles early studies, in bioinformatics several research groups to implement a new algorithm I had an impact. It reads based solely on Illumina sequence of short sequence that led to the creation of several projects of this genome, the length of 52 nucleotide sequence only the installation of the genome of 2,25-Gb of giant panda has been completed from Illumina sequence recently Reads the average, field to discuss some of the practical aspects such as the submission of assembly data to public repositories and filtering of such data take into account recent developments.

Without the need for a sequence database, new sequencing peptide sequence de is derived from the mass spectrum. It is also the most important software modules peaks. Independent sequence database is the preferred method for investigating the biological unsequenced identification and de novo sequencing, the new peptide. Even if it can help de novo sequence database is to improve the identification of the database search based peptides minutes between consistency and de novo sequence database significantly peptide sequence, verify the accuracy of the identification to.