depending on the particular thermostable DNA polymerase used, the fidelity of the PCR may vary (Speksnijder et al. 2001). Although careful analysis of secondary interactions should identify discrepancies due to misincorporation of nucleotides during PCR, there is a danger that the presence of novel taxa may be assumed as a result of infidelity in DNA replication. Many prokaryotes have several rRNA operons and even though the rRNA coding regions are usually highly similar, they are not necessarily identical. The cloning step in rRNA analysis separates not only the rRNA genes of different organisms but also the different genes. Thus, slightly different gene fragments could originate from one strain and would not indicate the presence of closely related organisms (Farrelly et al. 1995). This is of concern when making conclusions about biodiversity from data obtained with rRNA gene clone libraries.

DOI link for depending on the particular thermostable DNA polymerase used, the fidelity of the PCR may vary (Speksnijder et al. 2001). Although careful analysis of secondary interactions should identify discrepancies due to misincorporation of nucleotides during PCR, there is a danger that the presence of novel taxa may be assumed as a result of infidelity in DNA replication. Many prokaryotes have several rRNA operons and even though the rRNA coding regions are usually highly similar, they are not necessarily identical. The cloning step in rRNA analysis separates not only the rRNA genes of different organisms but also the different genes. Thus, slightly different gene fragments could originate from one strain and would not indicate the presence of closely related organisms (Farrelly et al. 1995). This is of concern when making conclusions about biodiversity from data obtained with rRNA gene clone libraries.

depending on the particular thermostable DNA polymerase used, the fidelity of the PCR may vary (Speksnijder et al. 2001). Although careful analysis of secondary interactions should identify discrepancies due to misincorporation of nucleotides during PCR, there is a danger that the presence of novel taxa may be assumed as a result of infidelity in DNA replication. Many prokaryotes have several rRNA operons and even though the rRNA coding regions are usually highly similar, they are not necessarily identical. The cloning step in rRNA analysis separates not only the rRNA genes of different organisms but also the different genes. Thus, slightly different gene fragments could originate from one strain and would not indicate the presence of closely related organisms (Farrelly et al. 1995). This is of concern when making conclusions about biodiversity from data obtained with rRNA gene clone libraries.