摘要
通过对“一步法”RNA提取方法的改进,简化了提取程序,并保证了RNA的质量。根据马铃薯X病毒(PVX)基因序列设计合成了一对特异性引物,运用反转录PCR(RT-PCR)对PVX-RNA进行扩增,成功得到了与预期大小相一致的308 bp片段,而对照未得到任何产物,同时对反转录酶AMV用量做了不同的处理,得出AMV仅用1u仍能得到较好扩增的效果。从而建立了经济简便的PVX的RT-PCR检测体系,为PVX的防治、脱毒苗的检测提供有效手段。
Total RNA was extracted with an improved one-step method from tobacco leaves infected with potato virus X (PVX). The more quality of tested RNA was very good, which indicated that the improved method was very effective and simple. A pair of specific primers were designed and synthesized based on the homologous nucleotide sequence of different isolates of PVX by the reverse transcription and polymerase chain reaction (RT-PCR), a 308 bp DNA fragment was amplified successfully from PVX of Shandong isolate, and no fragment was obtained from control. With only 1U Reverse Transcriptase XL (AMV) , the amplified result were still very good. The results showed that an economical and convenient RT-PCR detection system of PVX was set up.
出处
《园艺学报》
CAS
CSCD
北大核心
2003年第6期687-689,共3页
Acta Horticulturae Sinica
关键词
马铃薯
X病毒
RT-PCR检测
反转录PCR
PVX (Potato virus X)
Virus
Detection
Reverse transcription and polymerase chain reaction (RT-PCR)
