Rule G S, Montagna R A, Durst R A
Analytical Chemistry Laboratories, Cornell University, Geneva, NY 14456-0462, USA.
Clin Chem. 1996 Aug;42(8 Pt 1):1206-9.
We describe a rapid method for visually determining specific DNA sequences at femtomole concentrations. Liposomes, encapsulating a red dye and labeled with oligonucleotide, were used in a capillary migration-sandwich hybridization assay. Capture probe was immobilized on nitrocellulose strips, and liposomes, migrating along each strip, formed a visually discernible band in the presence of target DNA. One femtomole of synthetic target sequence could be detected in < 10 min. Sufficiently stringent hybridization conditions can be used to allow the discrimination of a 10% mismatch sequence from perfectly complementary DNA. A 366-base PCR product was detected at 200 fmol.
我们描述了一种在飞摩尔浓度下目视确定特定DNA序列的快速方法。在毛细管迁移-夹心杂交分析中，使用了包裹红色染料并标记有寡核苷酸的脂质体。捕获探针固定在硝酸纤维素条上，沿着每条条迁移的脂质体在存在靶DNA的情况下形成可见的条带。在不到10分钟内可检测到1飞摩尔的合成靶序列。可以使用足够严格的杂交条件来区分10%错配序列与完全互补的DNA。在200飞摩尔时检测到了一个366碱基的PCR产物。
