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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

Size

Catalog number selected: AX5810

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PowerSeq™ CRM Nested System, Custom
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Massively Parallel Sequencing of Mitochondrial Control Region

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.

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.

PowerSeq™ CRM Nested System, Custom, simplified workflow

Adapted from Lange et al. (2014) BMC Genomics 15, 63.

Balanced Representation of Pooled Libraries

Representative balanced profile of PowerSeq™ CRM Nested System, Custom.

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

What's in the box?

Item Part # SizeConcentrationAvailable 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 × 25μl10ng/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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Certificate of Analysis

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Use Restrictions

Not For Medical Diagnostic Use.

Storage Conditions

EE

Patents and Disclaimers

U.S. Pat. No. 6,242,235, Australian Pat. No. 761757, Canadian Pat. No. 2,335,153 and other patents and patents pending.

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