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Third Generation Sequencing

PACBIO RS II

The PacBio RS II is a Single Molecule, Real-Time (SMRT®) DNA Sequencing System that provides the highest consensus accuracy and longest read lengths of any available sequencing technology.

SMRT technology based on the natural process of DNA replication: an active DNA polymerase, which immobilized at the bottom of each ZMW (zero-mode waveguides), attached itself to a strand of DNA to be replicated and examine the base at the point it is attached, which then identify the nucleotide and the whole sequence. In order to detect the base, each of the four nucleotides A, C, G and T are labeled with a different fluorescent dye having a distinct emission spectrum, enable real time observation of DNA synthesis.

SMRT Sequencing can use for de novo assembly, characterization of genetic variation, methylation analysis, microbiology studies, etc. It can also use for RNA Sequencing in order to study isoforms because of the extraordinarily long reads.

Advantages:

  • Extraordinarily long reads: Depending upon starting library, half of the data are in reads >14,000 base pairs long with the longest reads over 40,000 base pairs.

  • Extremely high accuracy: Perform de novo assembly of genomes and detect variants with greater than 99.999% accuracy. Sequence individual molecules with 99% accuracy at greater than Sanger lengths.

  • Exquisite sensitivity: Detect minor variants that are present at a frequency less than 0.1%.

  • Shortest run time: Sequence for as little as 30 minutes and still get reads longer than Sanger sequencing.

  • Least GC bias: Sequence through regions of extremely high or low GC content with ease, resulting in more even coverage.

  • No amplification bias: Samples need not be amplified, improving coverage uniformity and avoiding artifacts.

For more information, please find http://www.pacificbiosciences.com

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