Related Experiment Video
Updated: Apr 18, 2026
07:24
Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing
Published on: February 10, 2023
2.1K
A high-throughput next-generation sequencing assay for the mitochondrial genome
Shale Dames1, Karen Eilbeck, Rong Mao
1ARUP Laboratories, ARUP Institute for Clinical and Experimental Pathology, 500 Chipeta Way, Salt Lake City, UT, 84108, USA, shale.dames@aruplab.com.
Methods in Molecular Biology (Clifton, N.J.)
|January 30, 2015
Summary
Next-generation sequencing (NGS) effectively analyzes mitochondrial DNA (mtDNA) and detects heteroplasmy. This protocol details an mtDNA enrichment method for Illumina sequencing, with an alignment script available for download.
Area of Science:
- Genomics
- Molecular Biology
- Bioinformatics
Background:
- Mitochondrial genome (mtDNA) sequencing is crucial for understanding cellular respiration and disease.
- Detecting heteroplasmy, the presence of multiple mtDNA variants, requires sensitive and accurate methods.
Purpose of the Study:
- To present a comprehensive protocol for mitochondrial genome (mtDNA) enrichment.
- To enable efficient library preparation and sequencing of mtDNA using Illumina next-generation sequencing (NGS) platforms.
- To provide a supporting command-line script for data analysis.
Main Methods:
- Mitochondrial DNA (mtDNA) enrichment technique.
- Library preparation for Illumina sequencing.
- Next-generation sequencing (NGS) on Illumina platforms.
- Command-line alignment script for data processing.
Main Results:
- The protocol facilitates robust mtDNA enrichment.
- Successful library preparation and subsequent sequencing were achieved.
- An alignment script was developed for efficient data analysis.
Conclusions:
- This protocol provides a reliable method for mitochondrial genome (mtDNA) sequencing and heteroplasmy detection.
- The integration of enrichment, library preparation, and sequencing on Illumina platforms streamlines the process.
- The provided script aids in the downstream analysis of sequencing data.
Related Concept Videos
03:00Next-generation Sequencing
102.2K
The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features....
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features....
102.2K
03:21RNA-seq
12.7K
RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
12.7K
02:59Animal Mitochondrial Genetics
10.2K
Among all the organelles in an animal cell, only mitochondria have their own independent genomes. Animal mitochondrial DNA is a double-stranded, closed-circular molecule with around 20,000 base pairs. Mitochondrial DNA is unique in that one of its two strands, the heavy, or H, -strand is guanine rich, whereas the complementary strand is cytosine rich and called the light, or L, -strand. Compared to nuclear DNA, mitochondrial DNA has a very low percentage of non-coding regions and is marked by...
10.2K