DNA nanoball sequencing
DNA sequencing nanoballs has a high-throughput technology used for sequencing of the entire genome sequence of an organism. Complete genome of the company is using this technology to sequence samples that were submitted by independent researchers. Way is to use a rolling circle replication to amplify small fragments of genomic DNA to the DNA nanoballs. Then unlinked sequencing by ligation is used to determine the nucleotide sequence. This method of DNA sequencing, can be sequenced to implement DNA, the nanoballs numerous and at a lower cost compared to the next-generation sequencing platforms other drugs. However, it can be read to reference genome mapping of short it is difficult to be determined by each DNA nanoballs the short sequence of DNA. It has been used for a variety of the genome project, this technology has been planned to be used more.
DNA sequencing nanoballs, has a high-throughput sequencing technology used for the determination of the entire genome sequence of an organism. Way is to use a rolling circle replication to amplify small fragments of genomic DNA to the DNA nanoballs. Associated with additional DNA, and then fluorescence probe is connected to the relevant anchor sequence with known sequences in the DNA template. The main purpose, is determined by the associated probe fluorescence and tyrant. The method can be sequenced to run at a lower cost than the drug generation sequencing platforms other DNA nanoballs many DNA. However, limitations of this method is to produce a short sequence of only DNA has become a challenge to move the reading of the reference genome. The company has used the DNA sequence to sequence nano ball samples that were submitted from the complete genome researchers.
The ligated fragments circularizing fragments and adapter sequence, the fragment (BP) cleavage of a small 400-500 base pairs, DNA sequences nanoballs involves the isolation of DNA to be sequenced. As a result of the single-stranded copies of many of the fragments, fragment of circular, is copied by a rolling circle replication. Head, and I pushed into the DNA nanoballs to tail in a thread long DNA copy associate. In nanoballs, and adsorption to the input cell sequence. Unlinked sequencing reactions, contact the site of the nucleotide specific nanoballs by connecting a fluorescent probe DNA. Color fluorescence in each interrogation position is detected by the high-resolution camera. Is used to analyze the fluorescence data, the production of each call, as well as for applying the 35-BP mate pair, bioinformatics, reads the reference genome. Genome is assembled, were identified polymorphisms all present in the sequence.
The lysed cells were extracted from the cell lysate DNA. High molecular weight DNA, often for long, megabase pairs some destroyed sonicated double-stranded DNA at random intervals. DNA sample is most effective if it contains a narrow range of length reads the mapping sequencing already. Therefore, sequencing of DNA fragments and selection of fragment length ideal, are separated polyacrylamide gel electrophoresis, size by (PAGE). Was purified by gel extraction as a result of DNA having (400-500 base pairs, typically) length of a narrow range, the DNA of the appropriate size range.
Adapter DNA sequence must be applied to DNA fragments of the DNA sequences of unknown unknown DNA clearance, as is known. I was PCR amplified the DNA and, attached to the left and right sides of the DNA fragments in the first round of (AD1) adapter ligation of the left and right adapters. Left AD1 is modified to create a single-stranded termini additional are connected to one another so as to form a circular DNA. Were cut 13 BP right right adapter DNA, restriction enzymes, are added. This results in double-stranded DNA linear. Are connected to the ends of (Ad 2) linear DNA in the left and right sides of the adapter sequence, the product is amplified using PCR. It is modified so that after a sequence of Ad 2, connect to each other to form a circular DNA. Restriction enzyme is used again to cleave circular DNA left BP AD1 13. The results, linear DNA fragments are (Figure 3). The lateral coupling (AD3) adapter sequence to the left and right of the linear DNA, was amplified by PCR product. The adapter is fixed to couple to each other because they form a circular DNA. Cutting the DNA of 26 bp to the left of the AD3, 26 bp to the right ad 2 depending on the type III restriction EcoP15, were added. In this step, to remove most of the DNA, and the DNA linearized again. Was amplified by PCR to combine the left and right adapters (AD4) DNA, the product, AD4 sequence is modified such that they bind to each other. The result is a complete circular DNA template.