PowerSeq® CRM Nested System, Custom
Produce MPS Libraries Ready for Mitochondrial Control Region Analysis
- Massively parallel sequencing of mitochondrial HVI, HVII and HVIII control regions in a single reaction
- Nested amplification protocol greatly reduces number of steps and time required for library preparation
- Small amplicons ranging from 144–237bp for better results with compromised samples
Catalog Number:
Size
Catalog Number: AX5810
Massively parallel sequencing (MPS) allows laboratories access to mitochondrial DNA (mtDNA) analysis using a more sensitive and high-throughput workflow compared to traditional sequencing methods. Increased mixture deconvolution and heteroplasmy resolution are achieved by deep sequencing coverage and digital read counts. Additionally, the use of small amplicons to sequence the mitochondrial control region improves sequencing results from degraded samples.
The PowerSeq® CRM Nested System, Custom, generates 10 small amplicons covering the control region of the mitochondrial genome in one multiplex. The targeted regions for amplification are designed to be in a range of 144–237bp to ensure optimal results from degraded samples. The system uses a nested amplification protocol that greatly reduces the number of steps and time required to produce libraries ready for sequencing. The protocol consists of a single PCR step to both amplify the target amplicons and incorporate indexed sequencing adapters.
Sequencing data can be analyzed using GeneMarker HTS Software (SoftGenetics, LLC).
The PowerSeq® CRM Nested System is now approved for use in laboratories that generate DNA profiles for submission to the National DNA Index System (NDIS).
For more information, see: Brandhagen, M.D. et al. (2019) Validation of NGS for mitochondrial DNA casework at the FBI Laboratory. Forensic Sci. Int. Genet. 44, 102151.
Simplified Workflow Using Nested Amplification Protocol
PowerSeq® CRM Nested System, Custom, combines primers needed for amplifying the targeted sequences with primers that contain unique indexing and adapter sequences for sequencing on the MiSeq® instrument. This workflow greatly simplifies library preparation by saving time, decreasing sample loss and reducing data variability.
Adapted from Lange et al. (2014) BMC Genomics 15, 63.
Balanced Representation of Pooled Libraries
Representative balanced profile of PowerSeq® CRM Nested System, Custom. 2800M Control DNA (500pg; n=4) was amplified with the PowerSeq® CRM Nested System. A pool of 16 samples was sequenced simultaneously using MiSeq® Reagent Kit v3. Balance data was analyzed with ExactID® (Battelle Memorial Institute).
Specifications
Catalog Number:
What's in the box?
| Item | Código | Tamanho | Concentration | Available Separately |
|---|---|---|---|---|
PowerSeq® CRM Nested 10X Primer Pair Mix |
AX579A | 1 × 250μl | ||
PowerSeq® Nested System Index Primer 1 D701 |
AX580A | 1 × 75μl | ||
PowerSeq® Nested System Index Primer 1 D702 |
AX582A | 1 × 75μl | ||
PowerSeq® Nested System Index Primer 1 D703 |
AX583A | 1 × 75μl | ||
PowerSeq® Nested System Index Primer 1 D704 |
AX584A | 1 × 75μl | ||
PowerSeq® Nested System Index Primer 1 D705 |
AX585A | 1 × 75μl | ||
PowerSeq® Nested System Index Primer 1 D706 |
AX586A | 1 × 75μl | ||
PowerSeq® Nested System Index Primer 1 D707 |
AX587A | 1 × 75μl | ||
PowerSeq® Nested System Index Primer 1 D708 |
AX588A | 1 × 75μl | ||
PowerSeq® Nested System Index Primer 1 D709 |
AX589A | 1 × 75μl | ||
PowerSeq® Nested System Index Primer 1 D710 |
AX590A | 1 × 75μl | ||
PowerSeq® Nested System Index Primer 1 D711 |
AX591A | 1 × 75μl | ||
PowerSeq® Nested System Index Primer 1 D712 |
AX592A | 1 × 75μl | ||
PowerSeq® Nested System Index Primer 2 D501 |
AX593A | 1 × 75μl | ||
PowerSeq® Nested System Index Primer 2 D502 |
AX594A | 1 × 75μl | ||
PowerSeq® Nested System Index Primer 2 D503 |
AX595A | 1 × 75μl | ||
PowerSeq® Nested System Index Primer 2 D504 |
AX596A | 1 × 75μl | ||
PowerSeq® Nested System Index Primer 2 D505 |
AX597A | 1 × 75μl | ||
PowerSeq® Nested System Index Primer 2 D506 |
AX598A | 1 × 75μl | ||
PowerSeq® Nested System Index Primer 2 D507 |
AX599A | 1 × 75μl | ||
PowerSeq® Nested System Index Primer 2 D508 |
AX600A | 1 × 75μl | ||
|
2800M Control DNA |
DD710A | 1 × 25μl | 10ng/μl | View Product |
PowerSeq® 5X Master Mix |
DP116A | 1 × 500μl | ||
|
Water, Amplification Grade |
DW099A | 2 × 1,250μl | View Product | |
|
Proteinase K |
V302A | 1 × 10mg | View Product |
SDS
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Restrições de uso
Not For Medical Diagnostic Use.Condições de armazenamento
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