Localization In Situ of Specific mRNA Using Thymine‐Thymine Dimerized DNA Probes
Localization In Situ of Specific mRNA Using Thymine‐Thymine Dimerized DNA Probes
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DOI：
10.1111/j.1440-1827.1990.tb02492.x
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发表时间：
1990-11
影响因子：
2.2
通讯作者：
T. Koji;P. Nakane
中科院分区：
文献类型：
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作者：
T. Koji;P. Nakane
原位杂交已被认为是一种强大的技术，用于定位特定的核酸，如mRNA和病毒DNA在个别细胞。对于原位杂交，从分辨率、探针稳定性和人身安全的观点来看，使用非放射性探针被认为上级使用放射性探针。虽然目前有各种非放射性标记，但有些会干扰杂交体的形成，有些会增加空间位阻并阻止标记的核酸探针渗透到细胞和组织中。最近，我们发展了一种以胸腺嘧啶（TT）二聚DNA作为非放射性探针的方法。这很容易实现，因为它不需要将标记的DNA与未反应的标记化合物分离，并且T-T二聚体不会改变探针的化学和物理性质。在本文中，我们描述了适合TT二聚DNA探针的组织处理程序，以及最近在方法学方面的新发现，特别是使用合成的寡脱氧核苷酸探针。《病理学杂志》40：793 - 807，1990年。
In situ hybridization has been recognized as a powerful technique for localizing specific nucleic acids such as mRNA and viral DNA in individual cells. For in situ hybridization, the use of a non radioactive probe is considered superior to that of a radioactive one from viewpoints of resolution, probe stability and personal safety. Although various non‐radioactive labels are currently available, some interfere with the formation of hybrids and some increase steric hindrance and prevent penetration of the labeled nucleic acid probe into cells and tissues. Recently, we have developed a method using thymine thymine (TT) dimerized DNA as a nonradioactive probe. This is simple to make, since it dose not require separation of labeled DNA from unreacted labeling compounds, and T‐T dimer will not alter the chemical and physical nature of the probe. In this paper, we describe the tissue processing procedures that are suited for the TT dimerized DNA probes, and the recent new findings on methodological aspects, particularly the use of synthetic oligodeoxynucleotide probes. Acta Pathol Jpn 40: 793‐807, 1990.