1μg ChrMT m.1555 A>G Ref Std Hereditary Deafness

Product Overview

This standard contains a precisely calibrated homoplasmic point mutation at position m.1555 A>G in the mitochondrial 12S rRNA gene, one of the most common genetic causes of aminoglycoside-induced and non-syndromic hearing loss. Derived from characterized cell lines with confirmed mitochondrial genotype, it provides a gold-standard control for ensuring accurate detection of this clinically significant mutation in diagnostic laboratories .

Product Features

Authentic Mutation Profile

Contains the pathogenic m.1555 A>G mutation in homoplasmic form (>99% mutant allele frequency), verified by Sanger sequencing, pyrosequencing, and next-generation sequencing (>10,000x coverage). This ensures reliable validation of assay sensitivity for detecting this low-abundance mutation .

Mitochondrial DNA Integrity

Purified to contain >95% mtDNA with minimal nuclear DNA contamination (<5%), as verified by quantitative PCR targeting mitochondrial (ND1) and nuclear (β-actin) genes. The DNA maintains native mitochondrial genome structure with average fragment size of 16.5 kb .

Comprehensive Quality Controls

Each lot undergoes rigorous testing including:

• Sequence validation across the entire mitochondrial genome

• Quantification by digital PCR (5 x 10⁴ copies/μL)

• Assessment of DNA integrity by agarose gel electrophoresis

• Verification of homoplasmic mutation status by clone sequencing

Usage

Assay Calibration

Use at a standard input of 10 ng per reaction to establish baseline performance metrics for m.1555 A>G detection assays. Prepare serial dilutions (0.1-100 ng) to determine limit of detection and linearity range.

Method Validation

Incorporate into validation studies to:

• Verify mutation detection accuracy across platforms

• Establish assay specificity against wild-type mtDNA

• Monitor inter-run variability in mutation calling

• Validate bioinformatics pipelines for mtDNA variant analysis

Storage and Handling

Store lyophilized DNA at room temperature for up to 36 months. Reconstitute in 100 μL molecular biology-grade water to achieve a concentration of 10 ng/μL. Store reconstituted aliquots at -20°C for up to 12 months, avoiding repeated freeze-thaw cycles .

FAQ

What makes the m.1555 A>G mutation clinically significant?

This mutation predisposes carriers to both non-syndromic hearing loss and severe, irreversible hearing loss following aminoglycoside antibiotic exposure, making accurate detection critical for personalized medicine and genetic counseling .

How does this standard differ from nuclear DNA controls?

As a dedicated mtDNA standard, it accounts for unique characteristics of mitochondrial genetics including heteroplasmy, maternal inheritance patterns, and high copy number, which are not addressed by nuclear DNA controls .

Can it be used to validate next-generation sequencing assays?

Yes, the standard is optimized for NGS validation, with uniform coverage across the mitochondrial genome enabling assessment of variant calling accuracy, coverage depth, and heteroplasmy detection limits .

Is wild-type control material available?

Yes, each lot includes a companion wild-type mtDNA standard from the same genetic background, enabling parallel validation of assay specificity and background noise levels .