Durrant I, Dacre B, Cunningham M
Amersham International, Buckinghamshire, UK.
Histochem J. 1995 Jan;27(1):89-93. doi: 10.1007/BF00164176.
Radioactive in situ hybridization predominantly utilizes either RNA probes or oligonucleotide probes. The properties of various formulations of [35S]UTP alpha S have been studied with respect to probe labelling and when applied to in situ hybridization. A new formulation has been prepared that combines a high physical concentration with a high specific activity so that, theoretically, predominantly full-length, high-specific-activity primary transcripts are produced. [alpha-33P]UTP can also be used for in situ hybridization and it compares favourably to 35S-labelled probes in terms of resolution and sensitivity. Both radiolabels can also be used to label oligonucleotide probes by a tailing reaction. [35S]dATP alpha S has been reformulated specifically for labelling oligonucleotides. The traditional stabilizer, dithiothreitol, which may cause precipitation within the tailing reaction buffer, has been replaced with an alternative stabilizer that avoids this problem while maintaining the stability of the nucleotide.
放射性原位杂交主要使用RNA探针或寡核苷酸探针。针对探针标记以及应用于原位杂交时，研究了[35S]UTPαS各种制剂的特性。已经制备了一种新的制剂，它结合了高物理浓度和高比活性，因此，理论上主要产生全长、高比活性的初级转录本。[α-33P]UTP也可用于原位杂交，在分辨率和灵敏度方面，它与35S标记的探针相比具有优势。两种放射性标记物也可通过加尾反应用于标记寡核苷酸探针。[35S]dATPαS已专门重新配制用于标记寡核苷酸。传统的稳定剂二硫苏糖醇可能会在加尾反应缓冲液中引起沉淀，已被另一种稳定剂取代，该稳定剂避免了这个问题，同时保持了核苷酸的稳定性。
