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Service

This section provides basic information about Service

De Novo Sequencing

Overview

De Novo sequencing sequences the whole genome sequence of unidentified microorganisms or animals and plants, and genetic factors of sarcoma and disorders can be discovered from the obtained results. By utilizing both the short read from Illumina Hiseq and long read from DNA Link’s Pacbio RS II, we provide service optimized for genome assemble finishing.

PacBio RS II System Read length distribution(P6-C4 lasted chemistry)

Benefit

In De Novo assemble, long reads have an advantage of creating long contigs from having many long read composition, and Pacific Biosciences SMRT (Single Molecule Real Time) RS, which is a third generation sequencer, also has an advantage of ability to read at length a single molecule in real time without the amplification step. Long reads allow to overcome repeat and GC rich region, and obtain single contig. In addition, it is also possible to obtain an optimal outcome by performing a hybrid assembly between long reads and short reads and adjusting error and accuracy accordingly.

  • scaffolding of large genomes
  • Longest read lengths with over 20,000 bp observed
  • Greater than 99.999% (QV 50) accurate sequencing
  • Sensitivity to detect minor variants at frequency less than 0.1%
  • No amplification bias and least impacted by GC content
  • Coverage uniformity
  • Shortest run time
  • De Novo Assembly: Reduce Ambiguities
  • Complete microbial genomes and improve assemblies of larger organisms
  • Read lengths up to 20kb, unbiased genome coverage, and high accuracy

Workflow

gDNA -> gDNA QC -> DNA Library Preparation -> On-Instrument DNA sequencing -> Bioinformatics

Sample requirements

sample quantity required :
 -short-insert libraries: ≥3 µg
 -Large-insert libraries: ≥20 µg
sample concentration :
 -short-insert libraries: ≥ 40ng/ul
 -Large-insert libraries: ≥ 200ng/ul
sample quality : A260/280:1.8-2.0, A260/230: ≥ 1.8 , Intacted DNA, RNA-free DNA, non-degraded

Turnaround time

About 4 weeks