Topic: DesignConstruction

Last updated: May 3, 2019

The development of single cell RNA sequencing or new generation sequencing has already replaced microarray technology, which used to be the way to analyze gene expression into multiple sections or regions of a genome.

It offers a number of advantages over microarray technologies; lower background noise, dynamic range in the quantification of gene expression level, and its capability to identify different isoforms. This technology has shown a stunning progress during the last decade in 2005 the amount of data produced by a Genome Analyzer could roughly reach one gigabase of Data, compared to 1.8 terabases on a single run in an equipment in 2014. Furthermore, the cost and efficiency of such studies has also been reduced in an enormous way, going from approximately three billion dollars in 2001 and 15 years to gather the whole sequence of a human genome to just one thousand dollars in 2014 and the capability to get up to 45 human genomes in a single day, at this rate we could be able to identify the genes responsible for most if not all known diseases and the conditions that may cause autism, cancer, schizophrenia, or heart disease, among others 5.The process followed previous to the use of sequencer technologies starts by choosing two or more samples of tissue in different conditions (occasionally natural and unhealthy), the RNA of these samples should then be isolated and pass through a library construction process, that involves the fragmentation of the DNA and cloning into vectors to posteriorly select the fragments with the most appropriate range size and adding links to the ends of those cDNA fragments. The fragments can later be analyzed in flow cell technology to obtain the nucleotides sequences of the cell samples.

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It has recently become a big challenge to find ways for sequencing DNA in a more and more rapid and cost effective manner. Therefore, the genetics and biology fields have become subjects of great attention and have led to a deeper understanding of life processes within molecular events. The enormous steps and advances reached on the evolution of technology used in the sequencing of DNA has allowed to find new applications and opportunities, but also has created new challenges for the processes that come before and after the processing of such massive amounts of data. In order to simplify the processing, analysis and increase the efficiency of current results, maintenance of long read sequences under a high and accurate quality must be achieved.


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