CAJ | 학술논문

目的介绍一种鸡胚原位杂交探针制备方法。方法携带Bmp4和Cath1基因特异片段的质粒载体，通过质粒小提试剂盒扩增，限制性内切酶线性化，酚氯仿抽提纯化， T3 RNA聚合酶体外转录，合成地高辛标记的反义探针，通过电泳及鸡胚原位杂交检测探针的质量。结果获得了效果良好的Bmp4和Cath1的RNA探针。结论用质粒小提试剂盒对质粒载体扩增，进而通过线性化、纯化、转录等步骤来制备原位杂交探针的方法，具有时间短、费用低、质量高、杂交效果好等优点。
목적개소일충계배원위잡교탐침제비방법。방법휴대Bmp4화Cath1기인특이편단적질립재체，통과질립소제시제합확증，한제성내절매선성화，분록방추제순화， T3 RNA취합매체외전록，합성지고신표기적반의탐침，통과전영급계배원위잡교검측탐침적질량。결과획득료효과량호적Bmp4화Cath1적RNA탐침。결론용질립소제시제합대질립재체확증，진이통과선성화、순화、전록등보취래제비원위잡교탐침적방법，구유시간단、비용저、질량고、잡교효과호등우점。
Objective To report a method for preparation of RNA probes for detection of Chicken Bmp4 and Cath1 in si-tu hybridization. Method Plasmids carrying specific sequences of Bmp4 and Cath1 genes were amplified using the Plasmid Mini Kit, linearized by restriction enzymes, extracted and purified by phenol-chloroform, and then synthesized to form anti-sense probes with T3 RNA polymerase in vitro. Quality of the prepared probes was tested by agarose gel electrophoresis and in situ hybridization in chicken. Results Well purified Bmp4 and Cath1 RNA probes with satisfactory results in in situ hybrid-ization were confirmed. Conclusions The reported method provides an excellent way to prepare RNA probes for chicken Bmp4 and Cath1 for in situ hybridization, which is economic, effective, with great results and excellent quality.