Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland.
J Basic Microbiol. 2012 Feb;52(1):104-9. doi: 10.1002/jobm.201000117. Epub 2011 Jun 9.
Analysis of 16S rRNA sequence diversity is widely performed for characterizing the biodiversity of microbial samples. The number of determined sequences has a considerable impact on complete results. Although the cost of mass sequencing is decreasing, it is often still too high for individual projects. We applied the multi-temperature single-strand conformational polymorphism (MSSCP) method to decrease the number of analysed sequences. This was a novel application of this method. As a control, the same sample was analysed using random sequencing. In this paper, we adapted the MSSCP technique for screening of unique sequences of the 16S rRNA gene library and bacterial strains isolated from biofilms growing on the walls of an ancient gold mine in Poland and determined whether the results obtained by both methods differed and whether random sequencing could be replaced by MSSCP. Although it was biased towards the detection of rare sequences in the samples, the qualitative results of MSSCP were not different than those of random sequencing. Unambiguous discrimination of unique clones and strains creates an opportunity to effectively estimate the biodiversity of natural communities, especially in populations which are numerous but species poor.
对 16S rRNA 序列多样性的分析被广泛用于描述微生物样本的生物多样性。确定的序列数量对完整的结果有很大的影响。尽管大规模测序的成本在降低，但对于单个项目来说，它通常仍然太高。我们应用多温度单链构象多态性（MSSCP）方法来减少分析的序列数量。这是该方法的一种新应用。作为对照，我们还使用随机测序方法对同一样本进行了分析。在本文中，我们适应了 MSSCP 技术，用于筛选从波兰一个古老金矿墙壁上生长的生物膜中分离出的细菌的 16S rRNA 基因文库的独特序列，并确定这两种方法的结果是否存在差异，以及随机测序是否可以替代 MSSCP。尽管它偏向于检测样品中罕见的序列，但 MSSCP 的定性结果与随机测序没有不同。对独特克隆和菌株的明确区分创造了有效估计自然群落生物多样性的机会，特别是在数量众多但物种贫乏的种群中。
