【摘要】 目的:建立一种快速、简便、高效的鸡胚整胚原位杂交地高辛标记的RNA探针的制备方法。方法:应用RT-PCR方法从鸡胚总RNA中扩增目的片段,连接到PGM-T克隆载体上,分别利用PGM-T载体中的SP6和T7RNA聚合酶上的启动子,以线性化的DNA为模板体外转录成地高辛标记的正反义RNA探针。结果:成功得到地高辛标记的RNA探针,探针在鸡整胚原位杂交中有杂交信号。结论:成功转录得到地高辛标记的正反义探针,为后续的以鸡胚为模型整胚原位杂交试验做好了探针的准备。更多还原

【Abstract】 Objective:To develop a fast,simple and efficient protocol for whole-mount in situ hybridization of chick embryos with digoxigenin(DIG)-Labeled RNA probe.Methods:The target fragment of gene was obtained by RT-PCR through total RNA of chicken,and then using the RNA polymerase promoter,based on linear DNA template in vitro transcribed into DIG-Labeled RNA probe.Results:Obtained the DIG-Labeled RNA probe and positive signals were viewed in chick embryos.Conclusion:The successful preparation of DIG-Labeled RNA probes lays an experimental foundation for whole-mount in situ hybridizatation of chick embryos which be used as an animal mode.更多还原