Polsky-Cynkin R, Parsons G H, Allerdt L, Landes G, Davis G, Rashtchian A
Clin Chem. 1985 Sep;31(9):1438-43.
Cloned Salmonella DNA, which has been immobilized irreversibly on plastic and agarose solid supports, can form hybrids in both single-layer and "sandwich" hybridization protocols. In single-layer hybridization, 3 micrograms of immobilized DNA bound at least 30 fmol of a specific 800-base DNA sequence (equivalent to 8.5 ng, or the amount of that sequence present in 4 X 10(10) organisms). In a 4-h sandwich hybridization protocol, as little as 14 amol (equivalent to 8 pg, or the amount of that sequence present in 1 X 10(7) organisms) of a 1600-base sequence of DNA could be detected. The methods described should be applicable to use with any set of probes--not just from Salmonella--that fulfill the criteria specified. The ability to perform DNA hybridizations on solid-phase matrices such as those used for immunoassay should bring DNA hybridization into the realm of routine clinical laboratory procedures.
已不可逆地固定在塑料和琼脂糖固体支持物上的克隆沙门氏菌DNA，可在单层和“夹心”杂交方案中形成杂交体。在单层杂交中，3微克固定化DNA至少能结合30飞摩尔特定的800个碱基的DNA序列（相当于8.5纳克，或该序列在4×10¹⁰个生物体中的含量）。在4小时的夹心杂交方案中，能检测到低至14飞摩尔（相当于8皮克，或该序列在1×10⁷个生物体中的含量）的1600个碱基的DNA序列。所述方法应适用于任何符合指定标准的探针组——不仅仅是来自沙门氏菌的探针。在用于免疫测定的固相基质上进行DNA杂交的能力应能使DNA杂交进入常规临床实验室操作领域。
