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Updated: Oct 2, 2026

Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons
Published on: August 29, 2014
Long PCR-amplified rDNA for PCR-RFLP- and Rep-PCR-based approaches to recognize closely related microbial species
Desouky Abd-El-Haleem1, Alice C Layton, Gary S Sayler
1The Center for Environmental Biotechnology and Department of Microbiology, The University of Tennessee, Knoxville, TN 37996-1605, USA.
Abstract:
Long PCR was used to amplify a 5-kb fragment of the bacterial ribosomal operon (16S-intergenic spacer region (ISR)-23S) from several Ralstonia eutropha strains (16S rDNA sequence similarity: 97-99%). Due to the large product size, amplicons from the different strains could be distinguished using restriction enzyme fragment length polymorphisms (RFLP) and repetitive PCR analysis (Rep-PCR) with the primer 1492r. These methods may prove useful in differentiating other bacterial strains with highly similar 16S rDNA sequences.
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