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.

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).
Protocols

Complete Protocol

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

PDF (138 KB)

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

Download SDS

PDF (597 KB)

Certificate of Analysis

See all certificates for AX5810

Use Restrictions

Not For Medical Diagnostic Use.

Storage Conditions

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.

Massively Parallel Sequencing of Mitochondrial Control Region

Balanced Representation of Pooled Libraries

Protocols

Complete Protocol

Specifications

What's in the box?

Certificate of Analysis

Use Restrictions

Storage Conditions

Patents and Disclaimers

Filters

Promega

PowerSeq™ CRM Nested System, Custom

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