NextSeq 2000
Mid-throughput sequencing for maximum flexibility
Available Kits - XLEAP-SBS Chemistry only.
Each kit is defined by the number of reads (single-end or clusters) and read length (determined by cycle count).
e.g. P4, 300 cycle flowcell will deliver about 1.8 billion single-end reads sequenced with the following read structure: 150-10-10-150
Sample Volume Requirements
- All flow cells (P1, P2, P3, P4): 60 µl (min 30 µl)
Library Concentration
Submit libraries at 2 nM. CTG will dilute the pool to the final loading concentration in RSB (provided by Illumina), which must be specified in your iLab submission. Loading concentration refers to the final concentration of library loaded onto an instrument for sequencing. See Illumina protocol for details.
Library Quality & Quantification
- Clean up libraries thoroughly to remove adapter/primer dimers and partial constructs.
- Use a Bioanalyzer, TapeStation or Fragment Analyzer to check fragment size and purity.
- Quantify libraries using qPCR for the most accurate measurement of usable fragments.
- Qubit can be used, but may include unwanted DNA (e.g., adapter dimers).
Sample Sheet Submission
A sample sheet (.csv) must be submitted with your library pool. This tells us how to demultiplex your data into FASTQ files. Use the Illumina template provided in iLab. Incorrect formatting may delay your run. Use letters (a-z) and numbers (0-9) only. No spaces, periods, underscores or hyphens.
Include the following fields:
- Sample_ID: Sample name
- Index: i7 sequence (forward)
- Index2: i5 sequence (forward)
- Sample_Project: iLab project number
Index Sequences Refer to your index kit documentation for correct sequences.
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