National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan.
Anal Biochem. 2011 Apr 1;411(1):152-4. doi: 10.1016/j.ab.2010.12.018. Epub 2010 Dec 13.
To rapidly detect specific genes, crude extracts prepared from rice seed grains were used as templates for PCR, the PCR products were digested with restriction enzymes or urasil-DNA glycosylase, and then matrix-assisted laser desorption ionization mass spectrometry (MALDI-TOF MS) was used to detect amplified DNA. It was possible to amplify small DNA fragments (50-60bp), but not large ones (>200bp), using crude extracts as the PCR template. This method can be completed within 1h, including extractions, and is well suited to automation for high-throughput analyses.
为了快速检测特定基因,使用从水稻种子中制备的粗提物作为 PCR 的模板,PCR 产物用限制酶或尿嘧啶-DNA 糖基化酶消化,然后用基质辅助激光解吸电离飞行时间质谱 (MALDI-TOF MS) 检测扩增的 DNA。使用粗提物作为 PCR 模板可以扩增小的 DNA 片段(50-60bp),但不能扩增大的片段(>200bp)。该方法可以在 1 小时内完成,包括提取过程,非常适合自动化高通量分析。