PCR-generated artefact from 16S rRNA gene-specific primers.
Artefacts consisting of concatenated oligonucleotide primer sequences were generated during sub-optimally performing polymerase chain reaction amplification of bacterial 16S rRNA genes using a commonly employed primer pair. These artefacts were observed during amplification for terminal restriction fragment length polymorphism analyses of complex microbial communities, and after amplification from DNA from a microbial culture. Similar repetitive motifs were found in gene sequences deposited in GenBank. The artefact can be avoided by using different primers for the amplification reaction.
Osborne
Catherine A
CA
Department of Microbiology and Immunology, University of Melbourne, Parkville, Victoria 3010, Australia.
Galic
Maja
M
Sangwan
Parveen
P
Janssen
Peter H
PH
D000818:Animals; D016477:Artifacts; D017931:DNA Primers; D007082:Ileum / Q000382:microbiology; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D012680:Sensitivity and Specificity; D012988:Soil Microbiology
D017931:DNA Primers; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S