Related Experiment Video
Updated: Jun 27, 2025

Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons
Published on: August 29, 2014
ddRAD Sequencing and DNA Barcoding
Vladislav Ivanov1, Kyung Min Lee2, Marko Mutanen1
1Ecology and Genetics Research Unit, University of Oulu, Oulu, Finland.
Abstract:
Double-digest restriction site-associated DNA sequencing is a library preparation protocol that enables capturing variable sites across the genome including single-nucleotide polymorphisms (SNPs). These SNPs can be utilized to gain evolutionary insights into patterns observed in DNA barcodes, to infer population structure and phylogenies, to detect gene flow and introgression, and to perform species delimitation analyses. The protocol includes chemically shearing genomic DNA with restriction enzymes, unique tagging, size selection, and amplification of the resulting DNA fragments. Here we provide a detailed description of each step of the protocol, as well as information on essential equipment and common issues encountered during laboratory work.
Related Concept Videos
Sanger Sequencing
Next-generation Sequencing
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features....
RNA-seq
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
Evolutionary Relationships through Genome Comparisons
Maxam-Gilbert Sequencing
Challenges of the Maxam-Gilbert Method
The...

