PowerSeq™ CRM Nested System, Custom

by Promega
Available SKUs: AX5810 |
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Description

  • Overview
  • Protocols
  • Specifications
  • 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 # Size 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

    Certificate of Analysis

    See all certificates for AX5810

    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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