Konat G W
Department of Anatomy, West Virginia University School of Medicine, Morgantown 26506-9128, USA.
Scanning Microsc Suppl. 1996;10:57-60.
The polymerase chain reaction (PCR) methodology can be employed to produce DNA hybridization probes. The major advantages of this paradigm over other techniques include superior specific activity of the probes, the versatility of sequence selection, the ability to produce short probes, and the simplicity of the procedure. We have further improved the efficiency of PCR probes by generating single stranded (ssDNA) probes that do not reanneal with themselves in solution, and hence, their availability for the interaction with the complementary sequences of the target is profoundly increased. Protocols for 32P-dCTP labeled and digoxigenin-dUTP labeled probes have been elaborated to maximize the incorporation rate of the label as well as to provide for the production of full-length probes. The ssDNA probes may be particularly suitable for nucleic acid detection in tissues by in situ hybridization.
聚合酶链反应（PCR）方法可用于生产DNA杂交探针。与其他技术相比，这种方法的主要优点包括探针具有更高的比活性、序列选择的通用性、能够生产短探针以及操作过程简单。我们通过生成在溶液中不会自身重新退火的单链（ssDNA）探针，进一步提高了PCR探针的效率，因此，它们与靶标互补序列相互作用的可用性大大增加。已经制定了32P-dCTP标记和地高辛配基-dUTP标记探针的方案，以最大限度地提高标记的掺入率，并确保生产全长探针。ssDNA探针可能特别适合通过原位杂交在组织中进行核酸检测。
