General Description
NextSeq 500 Illumina is a benchtop and lab-scale Sequencer from Illumina. It works in two configurations High and Mid Output and can sequence up to 400 M single reads of maximal length 300 bp. BaseSpace onsite Server is used for automated base calling and Fastq files generation.
NextSeq 500 Performance parameters | |||
Flow cell configuration | Read length | Output | Run time |
High Output (up to 400 M single reads) | 2 x 150 bp 2 x 75 bp 1 x 75 bp | 100-120 Gb 50-60 Gb 25-30 Gb | 29 h 18 h 11 h |
Mid Output (up to 130 M single reads) | 2 x 150 bp 2 x 75 bp | 32-39 Gb 16-19 Gb | 26 h 15 h |
Key Applications
- Transcriptome Sequencing (total RNA-Seq, mRNA-Seq, gene expression profiling)
- DNA-Protein Interaction Analysis (ChIP-seq)
- Small Whole-Genome Sequencing (microbe, virus)
- Targeted Gene Sequencing (amplicon-based, gene panel)
- Exome Sequencing
Sample Preparation
ü RNA and DNA isolation
- RNA and DNA to library preparation for sequencing can be isolated with TRIzol reagent or column-based methods with a purity as A260/280 = 1.8 – 2.0 measure by Nanodrop and high integrity (RIN ≥ 6.0) measured by Bioanalyzer or TapeStations.[KS2]
ü Library preparation
- DNA or RNA library can be prepared with home-made or commercially available kits offered by many companies compatible with Illumina technology. (link: protocols in NGS lab).
Further reading
- Intro_NextSeq_System_manual
- Illumina Adapter Sequences
- Index Adapters Pooling Guide
- ENCODE sequencing recommendations