摘要
①目的 探讨利用化学发光地高辛标记法制备牙龈卟啉菌 47A 1的全染色体探针的方法并测定标记效率。②方法 采用苯酚 氯仿 溴化十六烷基三甲胺 (CTAB)法制备牙龈卟啉菌 47A 1基因组DNA ,随机引物地高辛标记法制备核酸探针 ,在尼龙膜上直接点样 ,化学发光法检测。③结果 CTAB法可获得高质量的牙龈卟啉菌 47A 1染色体DNA ,地高辛标记 0 .3μg的牙龈卟啉菌核酸可获得 10ng探针DNA ,化学发光法检测最低可测出0 .0 0 2pg标记探针的存在。
Objective\ To investigate how the whole genomic DNA probe of Porphyromonas gingivalis to be prepared and determine the efficiency of labelling by digoxigenin. \ Methods\ Total cellular DNA from Porphyromonas gingivalis 47A 1 was extracted with CTAB method involving lysis of cells with SDS in the presence of proteinase K, treated with hexadecyltrimethyl ammonium bromide (CTAB) and phenol chloroform isopropanol. The purified DNA was digoxigenin labelled by a random primer technique using the DIG DNA Labelling Kit as the whole genomic DNA probe of Porphyromonas gingivalis.\ Results\ CTAB method produced high quality genomic DNA from Porphyromonas gingivalis. It could make 10ng probe from 0.3μg Porphyromonas gingivalis DNA by DIG labelling and as low as 0.002pg labelled DNA on nylon membrane can be detected. \ Conclusion\ It is a good way to get Porphyromonas gingivalis whole genomic DNA probe by random primer technique using the digoxigenin labelling.
出处
《青岛大学医学院学报》
CAS
2001年第3期173-175,共3页
Acta Academiae Medicinae Qingdao Universitatis
基金
国家自然科学基金资助项目 ( 3930 0 14 6 )
关键词
牙周炎
牙龈卟啉菌
DNA探针
标记
地高辛
periodontitis
porphyromonas gingivalis
DNA probe
markers
digoxin
