Single-stranded conformational polymorphism for separation of mixed rRNAS (rRNA-SSCP), a new method for profiling microbial communities

Barbara J. MacGregor, Rudolf Amann

Research output: Contribution to journalArticlepeer-review

12 Scopus citations


We show that non-denaturing gel electrophoresis, or single-stranded conformational polymorphism (SSCP), can be used to separate mixtures of full-length rRNAs. Individual bands can then be excised for identification by RT-PCR and sequencing. This has the advantage over profiling methods such as DGGE and T-RFLP that no PCR amplification is involved prior to sequencing; thus, extraction biases aside, it should yield a quantitative picture of community composition in terms of ribosome content. To simplify banding patterns, RNA subsamples (e.g. bacterial 16S rRNA) can first be isolated by magnetic bead capture hybridization. Alternatively, oligonucleotide-directed ribonuclease H (RNase H) digestion can be used to identify bands of interest by running digested samples in parallel to undigested ones. We illustrate the use of this technique to identify a potentially predominant species in a hypersaline microbial mat. We anticipate that rRNA-SSCP will be useful for community profiling; for clone library construction by directed cloning of individual rRNAs; and for following incorporation of radiolabeled substrates at the species level, by gel autoradiography, without advance information or guesswork about which species might be active and abundant.

Original languageEnglish (US)
Pages (from-to)661-670
Number of pages10
JournalSystematic and Applied Microbiology
Issue number8
StatePublished - Dec 4 2006
Externally publishedYes

Bibliographical note

Copyright 2015 Elsevier B.V., All rights reserved.


  • 16S
  • 18S
  • Microbial community profiling
  • Non-denaturing polyacrylamide gel electrophoresis


Dive into the research topics of 'Single-stranded conformational polymorphism for separation of mixed rRNAS (rRNA-SSCP), a new method for profiling microbial communities'. Together they form a unique fingerprint.

Cite this