Identify the differences with Illumina HiSeq 2500 or Illumina NextSeq 500 or Illumina MiSeq technology.
Re-sequence whole genomes and identify genetic variations, single base mutations, insertions and deletions compared to the reference genome. Based on your study's aims and the size of the genome, Eurofins Genomics will address your re-sequencing project using either HiSeq 2500 or NextSeq 500 or MiSeq technology.
Re-sequencing with Illumina HiSeq 2500
The HiSeq 2500 is ideal for whole genome re-sequencing of large eukaryotic genomes with well-known references. The tremendous data outputs allow sequencing of 6 human genomes per run or tens of fungal genomes per channel.
Re-sequencing with Illumina NextSeq 500
The NextSeq 500 is ideal for whole genome re-sequencing of large eukaryotic genomes with well-known references. The fast, integrated, sample-to-results workflow enables many sequencing applications—including transcriptomes, exomes, and targeted panels—in a single run.
Resequencing with Illumina MiSeq
With reads of up to 300 bp, the MiSeq sequences up to 24 bacterial genomes in one single run. Delivering turnaround times of only 3-4 weeks for such projects is a walk in the park for the Illumina MiSeq. If you are looking for high quality and fast re-sequencing of bacterial genomes, the MiSeq is the technology of choice.
Re-sequencing project specifications
Re-sequencing data delivers crucial information for disease pathways analysis, breeding studies, production strain optimization or metabolic engineering.
Our re-sequencing service comprises the following steps:
- Fragmentation of the genomic DNA ligation of sequencing adaptors
- Preparation of 2 shotgun libraries (i.e. with 200 bp and 400 bp fragment size) per sample (upon request)
- Shotgun sequencing on HiSeq 2500 or NextSeq 500 or MiSeq to 30-50-fold sequence coverage
- Combination with sequencing of different Mate pair / LJD libraries (upon request)
- Mapping of the sequencing data onto a reference sequence using appropriate software (reference guided assembly)
- Genome comparison by SNP and InDels analysis
- Sequence reads are not present on the reference genome (e.g. due to phage insertions or plasmids) are automatically sorted and will be delivered
We accept NGS grade high quality gDNA, tissue samples for this Service.
- Tissue: minimum 3-5 gm of tissues fast frozen in liquid nitrogen or in RNA later shipped in dry ice to Eurofins facility.
- gDNA: Submit 10-20 µg (30-50 µg if Mate pair library is requested) of RNA-free genomic DNA (NGS grade) of high molecular weight >40kb, Nanodrop A260/280 ratio >1.8; A260/230=2.0-2.2; and at a concentration of ~50 ng/µl (dissolve in low TE buffer).
Sample Quality Control:
- RNA free NGS grade gDNA isolation will be carried out using commercially available DNA isolation kit.
- The qualification and quantification of gDNA will be attained by agarose gel electrophoresis and NanoDrop/Qubit fluorometer analysis, respectively.
- DIN value estimation will be carried out by Agilent 4200 Tape Station.
- Preparation of 2 shotgun sequencing libraries using illumina TruSeq Nano DNA kit.
- Long jumping distance library / Mate pair library can be prepared upon request with a bridge gap of 3kbp and 8 kbp.
- Prepared libraries QC will be carried out using Agilent Tape Station.
- Prepared libraries will be sequenced on Illumina NextSeq 500 platform with 2 x 150 bp v2 chemistry or HiSeq 2500 with 2 x 125 bp v4 chemistry.
- Sequence data (base call files or bcl files) generated from the sequencer are de-multiplexed if required and converted to FASTQ files.
- Raw data will be available for download as a compressed archive of FASTQ files for each sample.
- Raw data can be post processed upon request.
- Comprehensive compiled report and data set in Pen Drive or Hard Disk.
- 5-7 weeks after arrival of your samples and all necessary information. It depends upon data size, scope, technology selected, number of samples and complexity of the project. The TAT is offered expecting no biological or technical difficulties for processing of all project samples.