A platform for human whole exome sequencing (WES) using target enrichment and library preparation for next generation sequencing.
(*) For unextracted samples.
(**) Turnover times are not guaranteed, may change according to your project.
Accepted Sample Type | Recommended Amount | Shipment method |
---|---|---|
Whole Blood (Fresh or Frozen) | 650μL -1mL | Blue ice for fresh blood Dry ice for frozen blood |
Buccal Swab | Follow the kit manufacturer's instructions | |
Tissue | 30 - 50 mg | Dry ice |
FFPE Tissue |
|
Room temprature |
Cell Pellet |
|
Dry ice |
Saliva | 650μL-1mL | Dry ice or blue ice |
Genomic DNA (RNA-free) | 50μL, >20 ng/μL | Dry ice |
We check your samples upon arrival, however we still require our users to do their own QC steps before sending samples.
Please use fluorometric measurements (e.g. Qubit, Quant-it), not absorbance measurements (Nanodrop, spectrophotometer).
The DNA should be in water or EB buffer, of high quality, and ideally have an A260/280: 1.8- 2.0 and A260/230: 2.0 – 2.2. Please use non-EDTA based buffers or low EDTA-buffers.
If your samples are below the required thresholds, please get in touch.
Once your samples arrive at SZAOMICS, we start by performing a reception control step in which we make sure the samples meet our requirements.
If the samples fail this quality control step, we will contact you to discuss possible options. Should you choose to proceed with samples not fulfilling the criteria it will be at your own risk, NGI will only make one attempt at library preparation and if the library preparation does not work you will be charged to the for the library preparation regardless.
If the samples pass reception control, the samples will be queued for library preparation.
At SZAOMICS, we offer whole exome sequencing including library preparation and enrichment using the Illumina DNA Prep with Enrichment Kit.
The protocol for library preparation involves DNA fragmentation and ligation of Illumina adapters with sample-specific barcodes. After sample and library preparation, a hybridization-based target enrichment step is performed using probes that contain specific sequences that are designed to bind complementary to the exome DNA sequences.
We evaluate the concentration of the libraries using qPCR and look at the fragment size distribution of the library molecules using Bioanalyzer. Once the libraries have passed this QC step, they are normalized, pooled, and queued for sequencing.
In general, we have been successfully generating libraries that provide enough good quality reads for sequencing of genomic DNA. However, the sequencing results from each library prep will d¬epend on the characteristics of the sample: