Before submitting your samples, please make sure you have completed the following steps:
1. Complete the online training to ensure each lab’s internal quality control and feedback.
3. Prepare sequencing libraries. Pool the barcoded samples if using multiplexed libraries. (See Library Prep.)
4. Check the quality of your samples (Qubit, qPCR and BioAnalyzer). (See Library QC.)
- Correct sample quantification is extremely important.
- Your Qubit and qPCR quantification should be largely consistent. If they are not, we suggest using the higher concentration to avoid potential over-clustering, which may significantly affect the data quality and yield.
- Double-check your pooled library before sending to us to avoid unintentional errors during sample pooling.
5. Provide pooled library as the following:
- 10 nM, 20 ul is recommended
- Minimum amount: 4 nM, 20 ul
We will set up the sequencing run in BaseSpace on your behalf. (See Sequencing.)
Please note: Minimum sequencing unit is one flow cell ( 300~400 million reads, which equal to the reads from two lanes in a 8-lane flow-cell used in most facilities). We do not accept libraries with 5′ inline barcodes, because Illumina sequencing is sensitive to constant sequence within the first few bases of the read. We use Illumina NextSeq 500 High Output kits (#FC-404-1004; FC-404-1002; FC-404-1005) and Medium Output kits (#FC-404-1003; FC-404-1001) to perform sequencing. Please make sure that your library preparation method is compatible with Illumina sequencing primers.