VALIDATED PANEL FOR DETECTION OF THE 24 HOTSPOTS FOR CARDIOVASCULAR DISEASE WITH LIBRARY PREP REAGENTS & BARCODES FOR ILLUMINA® SEQUENCING
NEXTFLEX® Cardiovascular Disease Amplicon Panel
- 100% coverage of 24 hotspots for cardiovascular disease
- 100% uniformity of amplicons on 0.2x coverage
- 384 unique barcodes available upon request
- Complete solution for targeted sequencing, including gene specific primers, PCR Master Mix, clean-up beads, and barcodes
- Incorporates a single-tube format
- Low input – Only 10 ng DNA isolated from fresh or frozen samples required
Coverage of 24 Hotspots for Cardiovascular Disease
The NEXTFLEX® Cardiovascular Disease Amplicon Panel contains primer pairs and reagents needed to amplify 24 hotspots for cardiovascular disease from DNA isolated from fresh or frozen material, and prepare libraries for Illumina® sequencing.
A Complete Solution for Sequencing of Cardiovascular Disease Hotspots
The NEXTFLEX® Cardiovascular Disease Amplicon Panel is a complete, validated solution that targets 24 hotspots for cardiovascular disease using 22 PCR primer pairs. This kit requires only 20 ng of DNA isolated from fresh or frozen samples. The convenient predesigned panel allows researchers to focus on data generation and analysis, not on labor-intensive primer design and target selection. This kit can be easily automated on liquid handlers, further simplifying workflow.
The NEXTFLEX® Cardiovascular Disease Amplicon Pane has the capability of multiplexing up to 1,700 samples at 100x coverage and 90 samples at 2000x coverage for detection of germline mutations on a single Illumina® 2×300 MiSeq® lane.
| HUGO Gene Name | Gene Name | Chromosome | reference SNP ID number |
|---|---|---|---|
| MTHFR | Methylenetetrahydrofolate reductase | chr1 | rs1801133 |
| MTHFR | Methylenetetrahydrofolate reductase | chr1 | rs1801131 |
| F5 | Factor V | chr1 | rs1800595 |
| F5 | Factor V | chr1 | rs6025 |
| F5 | Factor V | chr1 | rs118203906 |
| AGT | Angiotensin | chr1 | rs699 |
| APOB | Apolipoprotein B | chr2 | rs5742904 |
| AGTR1 | Angiotensin II receptor type 1 | chr3 | rs5186 |
| FGB | Fibrinogen | chr4 | rs1800790 |
| F13A1 | Coagulation factor XIII | chr6 | rs5985 |
| LTA | Lymphotoxin alpha | chr6 | rs1041981 |
| SERPINE1 | Plasminogen activator inhibitor type 1 | chr7 | rs1799889 |
| NOS3 | Nitric oxide synthase 3 | chr7 | rs2070744 |
| NOS3 | Nitric oxide synthase 3 | chr7 | rs1799983 |
| JAK-2 | Janus kinase 2 | chr9 | rs10974944 |
| F2 | Prothrombin F2 | chr11 | rs1799963 |
| ITGB3 | Integrin beta-3 | chr17 | rs5918 |
| ACE | Angiotensin I converting enzyme | chr17 | rs863223428* |
| APOE | Apolipoprotein E | chr19 | rs429358 |
| APOE | Apolipoprotein E | chr19 | rs7412 |
| PROCR | Protein C receptor | chr20 | rs867186 |
| PROCR | Protein C receptor | chr20 | rs9574 |
| CBS** | Cystathione β-synthase | chr21 | 844IN68 (no rs) |
* 283 bp Alu Insertion/Deletion targeted by this panel does not have dbSNP rs number. SNP rs863223428 is a different insertion which insertion point exactly coincides with 283 bp Alu I/D mutation.
** The amplicon for this gene also targets identical region in CBSL gene. CBS and CBSL encode proteins with identical amino acid sequence.
- NEXTFLEX® CVD Amplicon Primer Mix
- NEXTFLEX® PCR Enhancer 2
- NEXTFLEX® Hot Start PCR I Master Mix
- NEXTFLEX® Ligation Mix
- NEXTFLEX® Amplicon DNA Adapter
- NEXTFLEX® PCR II Barcoded Primer Mix
- NEXTFLEX® PCR II Master Mix
- Nuclease-free Water
- Resuspension Buffer
- NEXTFLEX® Cleanup Beads
REQUIRED MATERIALS NOT PROVIDED
- 10 – 50 ng of extracted genomic DNA (in up to 27 μL nuclease-free water)
- Ethanol 80% (room temperature)
- 96 well PCR Plate Non-skirted (Phenix Research™, Cat # MPS-499) or similar
- 96 well Library Storage and Pooling Plate (Fisher Scientific®, Cat # AB-0765) or similar
- Adhesive PCR Plate Seal (Bio-Rad®, Cat # MSB1001)
- Magnetic Stand -96 (Thermo Fisher® Scientific, Cat # AM10027) or similar
- Thermocycler
- 2, 10, 20, 200 and 1000 μL pipettes / multichannel pipettes
- Nuclease-free barrier pipette tips
- Microcentrifuge
- 1.5 mL nuclease-free microcentrifuge tubes
- Vortex
The NEXTFLEX® Cardiovascular Disease Amplicon Panel contains enough material to prepare 8, 48, or 96 samples for Illumina® sequencing from DNA isolated from fresh or frozen samples. The shelf life of all reagents is 12 months when stored properly. All components can be safely stored at -20°C.