The TruSight Cardio *.bed file lists all targets of enrichment for this assay. Filter by gene name (HGNC nomenclature) or reference the coordinates of your loci of interest in the human genome reference (hg19 build). See the product files for the gene list.
Visit the Safety Data Sheets (SDS) page and enter the applicable search terms.
Number of Samples
TruSight Cardio Sequencing Kit for MiSeq and MiSeqDx
MiSeq v2 chemistry, MiSeq reagent kit included
TruSight Cardio Sequencing Kit for NextSeq
NextSeq v2 chemistry, mid output
Expiration dates are printed on the kit box labels and reagent tubes. Illumina guarantees three months of shelf life from the time of shipment.
When sequencing on the NextSeq 550 System, you can pool 4 libraries containing 12 samples each (48 samples total). Make sure that each sample has a unique index combination.
Quantify the starting gDNA using a fluorometric-based method specific to double-stranded DNA (dsDNA). Run samples in triplicate to obtain more confident measurements. For details, see the input recommendations in the reference guide.
Sequence TruSight Cardio libraries using a paired-end 151-cycle run on the MiSeq, MiSeqDx (RUO), NextSeq 500, or NextSeq 550 Systems.
Due to the library sizes generated in TruSight Cardio, sequencing at longer read lengths increases the likelihood of sequencing into the adapter sequence.
When sequencing TruSight Cardio libraries on the MiSeq or MiSeqDx (RUO) with MiSeq v2 reagents, dilute libraries to a loading concentration of 8–12.5 pM. When sequencing on NextSeq 550 or NextSeq 500 with NextSeq v2 mid-output reagents, dilute to a loading concentration of 1.8 pM.
Results can vary based on the quantification method and instrument-to-instrument variability. For guidance, see the Cluster Optimization Overview Guide.
You can download the manifest file from TruSight Cardio Sequencing Kit Product Files.
Set up a paired-end, 151-cycle run on the MiSeq, MiSeqDx (RUO), NextSeq 500, or NextSeq 550 System.
Due to the library sizes generated in TruSight Cardio, sequencing at longer read lengths increases the likelihood of sequencing into the adapter sequence. See the TruSight Cardio Library Prep Reference Guide for information on pooling and dual-indexed sequencing.
To create subpanels, generate gene lists in HGNC nomenclature for the regions of interest. Upload this subpanel gene list when importing VCF files into VariantStudio, and then filter variants based on the shortened gene list.
For reference, see the complete TruSight Cardio gene list.
For sequencing of TruSight Cardio libraries on the MiSeq, use the MiSeq Reporter Enrichment workflow or the Generate FASTQ workflow. Illumina recommends preparing the sample sheet in the Illumina Experiment Manager (IEM). Libraries prepared with TruSight Cardio Sequencing kit (MiSeq, 12 samples) are dual-indexed, but only require a single i7 Index Read for demultiplexing. Set up the run for paired-end 151-cycle reads and a single 8 bp Index Read. FASTQ files can be uploaded to BaseSpace and analyzed using the BWA Enrichment App or the Isaac Enrichment App (v2.0 and v2.1 custom manifest workflow). Alternatively, use the MiSeq Reporter Enrichment workflow.
When using BaseSpace, set up the run in BaseSpace Prep tab. Select Nextera Rapid Capture from the dropdown menu under Library Prep kit to associate samples with the indexes provided in the TruSight Cardio Sequencing kit (NextSeq, 48 samples, Mid-output), E502, E503, E505, E506, and N701-12. Set up the run as a dual index paired-end 151-cycle sequencing run. FASTQ files streamed into BaseSpace can be analyzed using the BWA Enrichment App or the Issac Enrichment App (v2.0 and v2.1 custom manifest workflow).
When running the NextSeq in Standalone mode, enter the following parameters on the Run Setup Screen:
Read Type: Paired End
Read Length: Read 1: 151; Read 2: 151; Index 1: 8; Index 2: 8
When the run is complete, use the bcl2fastq2 converter for demultiplexing and FASTQ conversion. Create a sample sheet for demultiplexing using Illumina Experiment Manager. Choose NextSeq; Nextseq FASTQ only; Nextera Rapid Capture Enrichment Sample Prep kit. Perform data analysis using third-party software.