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 Delivery
- Sequencing pools delivered by Friday will be scheduled for sequencing the following week. The exact sequencing date is determined each Monday, and you will be notified of the scheduled date on the same day.
- Feel free to deliver your sequencing pool to our lab whenever it suits you, the lab is open all the time. Leave the pool in the black box inside the freezer which is located in lab D1426a and is marked for delivery of SeqOnly pools.
- Label the tubes clearly with the CTG project ID as generated in iLab.
- Upload the sample sheet in the corresponding project in iLab at the same time!
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.
More Info:
