Polyvinyl alcohol enhances detection of low abundance transcripts in early stage quail embryos in a nonradioactive whole mount in situ hybridization technique.
Polyvinyl alcohol enhances detection of low abundance transcripts in early stage quail embryos in a nonradioactive whole mount in situ hybridization technique.
复制标题
聚乙烯醇增强了非放射性整体原位杂交技术中早期鹌鹑胚胎中低丰度转录本的检测。
作者：
J. Barth;R. Ivarie
在鹌鹑早期胚胎整体原位杂交中，研究了聚乙烯醇（PVA）对吲哚硝基蓝四氮唑碱性磷酸酶显色反应的增强作用。以13-14体节对鹌鹑胚胎杂交中的生肌因子qmyoD检测为试验系统，10%的13-23 kDa和31-50 kDa PVA溶液最佳地增强了碱性磷酸酶显色反应。在增强的检测条件下，我们已经能够早在3体节对阶段测量qmyoD表达，而以前我们无法在10体节对阶段之前检测到qmyoD。体细胞qmyoD表达的PVA增强检测是特异性的，因为在与有义qmyoD探针的对照杂交中没有发现相应的信号。
We have tested the ability of polyvinyl alcohol (PVA) to enhance the indoxyl-nitro blue tetrazolium alkaline phosphatase color reaction in whole mount in situ hybridizations of early stage quail embryos. Using detection of the myogenic factor qmyoD in hybridizations of 13-14 somite-pair quail embryos as a test system, 10% solutions of 13-23 kDa and 31-50 kDa PVA optimally enhanced the alkaline phosphatase color reaction. With the enhanced detection conditions, we have been able to measure qmyoD expression as early as the 3 somite-pair stage, whereas previously we were unable to detect qmyoD prior to the 10 somite-pair stage. PVA-enhanced detection of somitic qmyoD expression was specific as no corresponding signal was found in control hybridizations with a sense qmyoD probe.