Whole Genome Sequencing on Sequel II

Nucleome Informatics is the only company in India to offer sequencing services on PacBio Sequel II sequencer. We offer Whole-genome sequencing service of Microbe, Plant and Animal Genomes. 

Sample Requirement

  1. Ten (CLR) to fifteen (CCS) ug of HMW gDNA of more than 50Kb long without any impurity
  2. Low Input DNA library is alos available. Please check with the technical team before placing the order.
  3. You can avail our HMW gDNA prep service too if you wish to send the tissue sample

HMW gDNA and Library QC

  1. Qubit
  2. Bioanalyser
  3. FEMTO Pulse
  4. PFGE

HMW gDNA Shearing and Size selection

  1. Shearing on Megaruptor 3
  2. Size Selection on Blupippin

Library Insert Size

  1. De novo assembly HiFi CCS Library (15 to 20Kb)
  2. De novo assembly CLR Library (30 to 40Kb)
  3. De novo assembly for low DNA input (20Kb)
  4. Microbial de novo assembly CCS Library (10-15Kb)
  5. Variant Detection CCS library (15 – 20Kb)
  6. Structural variant detection CLR library (>15Kb)

Sequencing on Sequel II platform

  1. PacBio Raw Coverage:
    1. Diploid genome: 50 to 60x (CLR) or min 30x-50x Hifi CCS coverage
    2. Polyploid Genome: 30x per haplotype (CCS)
    3. Complex Genome with large repeats: 120X coverage (CCS or CLR)
    4. Microbial Genomes: 200x CCS HiFi Coverage to achieve at least 30x Unique Molecule coverage per microbial genome
    5. Variant Detection: More than 15X HiFi for a human genome
    6. Structural Variant Detection: 5 to 25x CLR coverage

Data analysis

We use various tools to assemble the genomes. We build contig level assembly with PacBio Sequel II data and scaffold the genome with Optical map and HiC data after multiple rounds of polishing, gap filling and curation. Assessment of assembly is done using assembly stats like Contig N50, Scaffold N50, No of Gaps, Base accuracy, BUSCO Score, RNA Seq mappability, Phase Block resolution and Chromosomal level continuity. We offer the following type of assembly and scaffolding services;

  1. De novo contig level mosaic assembly
  2. Haplotype Phased diploid genome Assembly
  3. Trio Binning assembly
  4. Microbial genome assembly
  5. Polyploid genome assembly
  6. Scaffolding with Optical mapping 
  7. Superscaffolding with HiC data
  8. Polishing and error correction
  9. Genome Quality Assessment 
  10. Assembly Curation of previously developed genome assembly

For any query and more information on our services, please contact our sales team at salessupport @ nucleomeinfo.com. 

Pacific Biosystems PacBio Sequel II product.
20190322_10290_PacBio_Sequel2.CR2
Megaruptor 3 at Nucleome

ISO Sequencing

Coming up shortly. Stay tuned

Coming soon..