单链DNA图片
许多应用领域要求使用单链DNA模板。 The immobilization of a double-stranded PCR product for preparation of two single-stranded DNA fragments is probably the most widely used protocol for Dynabeads M-280 Streptavidin. 首先对两个PCR引物之一进行生物素化,生成PCR产物。 The PCR product is immobilized onto the beads for separation of the biotinylated and non-biotinylated strands. 洗脱条件可以是加热或碱变性。 所有多余的引物、核酸和缓冲液组分均被去除,无需离心或沉淀。

链霉亲和素偶联的Dynabeads不会抑制酶的活性。 This enables further handling and manipulation of the bead-bound DNA directly on the solid-phase.

Both the immobilized and the eluted single-stranded DNA template can be used in downstream applications such as MALDI-MS (1), pyrosequencing technology (2) and SNP analysis (3), as well as single-strand conformation polymorphism (SSCP), solid-phase DNA sequencing (4,5), DNA chips & microarrays, allele-specific extension and primer extension. 单链DNA模板还可用于体外诱变、核酸酶S1定位 (6)、探针制备和标记 (7)、差减杂交 (8,9,10) 及其他多种分子技术。

部分参考文献

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  2. Pourmand N. et.al. (2002) 多元焦磷酸测序。 Nucleic Acids Res.30(7):e31.
  3. Lindblad-Toh K.et.al. (2000) 小鼠单核苷酸多态性的大规模研究和基因分型。 Nature Genetics. 24:381-386.
  4. Hultman T. et.al. (1989) 采用磁珠作为固相支持物对基因组和质粒DNA进行直接固相测序。 Nucl. Acids Res. 17(13):4937-4946.
  5. Hultman T. et.al. (1991) 体外扩增质粒DNA的双向固相测序。 BioTechniques 10(1):84-93.
  6. Dziembowski A. et.al. (2001) 利用固相S1核酸酶定位法分析RNA的3'和5'端。 Anal. Biochem. 294:87-89.
  7. Beulieu M. et.al. (2001) PCR候选区域错配扫描: adaptation to quantitative, high-throughput genotyping. Nucleic Acids Res. 29(5):1114-1124
  8. Hansen-Hagge TE. et.al. (2001) 利用新型连接反应介导的消减 (LIMES) 法鉴定哺乳动物cDNA和基因组DNA中的样本特异性的序列。 Nucl. Acids Res.29(4):e20.
  9. Pradel N. et.al. (2002) Genomic subtraction to identify and characterize sequences of Shiga toxin-producing Escherichia coli O91:H21. Appl. Env. Microbiol. 68(5):2316-2325.
  10. Laveder P. et.al. (2002) 自消减cDNA文库的两步法构建策略。 Nucleic Acids Res. 30(9):e38.