Journal
JOURNAL OF MICROBIOLOGY
Volume 50, Issue 6, Pages 1071-1074Publisher
MICROBIOLOGICAL SOCIETY KOREA
DOI: 10.1007/s12275-012-2642-z
Keywords
pyrosequencing; 16S rRNA gene; PCR artifacts; PCR cycle number; DNA polymerase
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Funding
- National Academy of Agricultural Science, Rural Development Administration, Republic of Korea [PJ907127032012]
- Rural Development Administration (RDA), Republic of Korea [PJ907127032012] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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The effects of PCR cycle number and DNA polymerase type on 16S rRNA gene pyrosequencing analysis were investigated using an artificially prepared bacterial community (mock community). The bacterial richness was overestimated at increased PCR cycle number mostly due to the occurence of chimeric sequences, and this was more serious with a DNA polymerase having proofreading activity than with Taq DNA polymerase. These results suggest that PCR cycle number must be kept as low as possible for accurate estimation of bacterial richness and that particular care must be taken when a DNA polymerase having proofreading activity is used.
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