INVIEW Microbiome 16S-Full-Length

Hybrid Assembly Bacteria

ONT GridION & Illumina NovaSeq

Hybrid assembly for bacteria combines 30x long-read data with 300x short-read accuracy to enhance consensus quality in homopolymer regions and methylation motifs. This method achieves the most precise and reliable bacterial genome assemblies, often resulting in single-contig replicons for the majority of individual genomes.

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Applications

De novo assembly of bacteria (< 7 Mb) and intrinsic plasmids (no reference sequence required)

Highlights

No minimum sample number required
Sequencing one using GridION and one sample using Illumina
Polishing: Enhancement of the assembly quality through polishing with Illumina data, resulting in a more refined genomic sequence.

>> Sample Submission

Product Specifications & Ordering

Starting material

We are working on an optimised process. Until this process is established we do need 2 samples shipped to us. Both have the same sample requirements:

Sample Type: Purified DNA
Purity (OD260/280): 1.8-2.0

Volume: > 20µL
Concentration: 30 ng/µl using a fluorescence-based method
Minimum Amount: 600 ng
Resuspension Buffer Water, EB, or low TE (<0.1 mM EDTA)
Format: barcode labelled 1.5 ml safe-lock tubes
Shipment Method: Room temperature

Specifications

1st sample

Library preparation (amplification-free, including minimal fragmentation); using v14 library prep chemistry
Sequencing using Oxford Nanopore GridION (flow cell R10.4.1), on average 30x genome coverage
Draft Assembly: Utilizing Nanopore sequencing data to create an initial draft assembly of your genomic material.

2nd sample

Illumina-compatible PCR-based library preparation using Unique Dual Indexing
Sequencing on Illumina NovaSeq 6000 or NovaSeq Xplus (2 x 150 bp), on average 300x genome coverage

Combined bioinformatics analysis

Assembly Quality Assessment: Thorough evaluation of the draft assembly to ensure accuracy and completeness.
Polishing: Enhancement of the assembly quality through polishing with Illumina data, resulting in a more refined genomic sequence.
Annotation: Detailed annotation of the polished assembly, identifying genes and other functional elements.

Deliverables

Raw sequencing FASTQ data
Assembled Contigs: Delivered in FASTA format, ready for downstream applications.
Analysis Report: An HTML report summarizing findings and insights from the assembly process.
Circular genome plot as a publication-ready PNG
Annotated genome sequence of all contigs in GENBANK format (import directly into your favorite software like Sanger’s Artemis, SnapGene, Geneious Prime, Benchling, SeqBuilder Pro etc.)
Annotated genome features in GFF3 format
Annotated feature table, including the nucleotide sequence of each feature
Nucleotide sequences of all gene sequences in FASTA format
Amino acid sequences of all annotated protein sequences in FASTA format

Sample Submission Guidelines

For the sample preparation please view "starting material"

For the ordering process please be aware that for now you need to place 2 separate products in your cart for ONT sequencing and Illumina sequencing.

1. ONT Sequencing

For your ONT Sequencing you need the following barcodes:

ONT free barcodes >>

When using these barcodes please submit your ONT samples using this order page:

Bacterial Genome Sequencing on ONT >>

2. Illumina Sequencing

For your Illumina sample you need the following barcodes:

NGS free barcodes >>

When submitting the sample please use the following order page to order the Illumina sequencing and polishing (please select the Bacteria Hybrid assembly product)

INVIEW Resequencing >>

We strongly recommend to use the same sample name for the Illumina and ONT part (discrimation happens via the barcode)

Please note that we are strongly working on a much more convenient solution for you!

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