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- Overview
- Protocols
- Specifications
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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.
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).
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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 AX5810Use Restrictions
Not For Medical Diagnostic Use.Storage Conditions
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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