Differential amplification of rRNA genes by polymerase chain reaction.
The polymerase chain reaction (PCR) is used widely to recover rRNA genes from naturally occurring communities for analysis of population constituents. We have found that this method can result in differential amplification of different rRNA genes. In particular, rDNAs of extremely thermophilic archaebacteria often cannot be amplified by the usual PCR methods. The addition of 5% (wt/vol) acetamide to a PCR mixture containing both archaebacterial and yeast DNA templates minimized nonspecific annealing of the primers and prevented preferential amplification of the yeast small-subunit rRNA genes.
D000081:Acetamides / Q000494:pharmacology; D001105:Archaea / Q000235:genetics; D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D005784:Gene Amplification / Q000235:genetics; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction; D012329:RNA, Bacterial / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013045:Species Specificity
D000081:Acetamides; D004269:DNA, Bacterial; D012329:RNA, Bacterial; C030686:acetamide