| Human, Bacterial and Fungal Amplicon Collection and Processing for Sequencing
|
|
Author:
Date:
2015-05-20
[Abstract] Sequencing taxonomic marker genes is a powerful tool to interrogate the composition of microbial communities. For example, bacterial and fungal community composition can be evaluated in parallel using the 16S ribosomal RNA gene for bacteria or the internal transcribed spacer region in fungi. These are conserved regions that are universal to a taxonomic clade, yet have undergone some degree of evolution such that different lineages can be differentiated. Conserved regions are used for design of universal priming sites that allow amplification of the marker gene out of a mixed microbial community. Here, we describe our standard operating procedure to collect and sequence 16S rRNA and ITS1 amplicons from human skin. We use the 16S rRNA V1-V3 region for skin samples, as it has greater power ...
[摘要] 测序分类标记基因是一个强有力的工具，以询问微生物群落的组成。 例如，可以使用细菌的16S核糖体RNA基因或真菌中的内部转录间隔区平行评估细菌和真菌群落组成。 这些是对分类学分支是通用的保守区，但已经经历了某种程度的进化，使得不同谱系可以分化。 保守区用于设计允许从混合微生物群落扩增标记基因的通用引发位点。 在这里，我们描述我们的标准操作程序收集和序列16S rRNA和ITS1扩增子从人类皮肤。 我们使用16S rRNA V1-V3区域的皮肤样本，因为它有更大的权力分类皮肤中的常见葡萄球菌。 该方案适用于扩增子的454焦磷酸测序。
|
|
|
| Library Construction for Genome-wide Bisulfite Sequencing in Plants
|
|
Author:
Date:
2013-12-20
[Abstract] DNA methylation is the most studied epigenetic modification, which involves the addition of a methyl group to the carbon-5 position of cytosine residues in DNA. DNA methylation is important for the regulation of gene expression. Bisulfite sequencing is the gold standard technique for determining genome-wide DNA methylation profiles in eukaryotes. This protocol describes how to prepare libraries of genomic DNA for whole-genome bisulfite sequencing in Arabidopsis, which could be adapted for use in other plant species.
[摘要] DNA甲基化是研究最多的表观遗传修饰，其涉及将甲基添加到DNA中胞嘧啶残基的碳-5位置。 DNA甲基化对于基因表达的调节是重要的。 亚硫酸氢盐测序是确定真核生物全基因组DNA甲基化谱的金标准技术。 该协议描述了如何在拟南芥中制备用于全基因组亚硫酸氢盐测序的基因组DNA文库，其可以适用于其他植物物种。
|
|
|