实验原理:根据颈环引物的空间结构来提高反转录的特异性,从而反转录特定的small RNA,在实时定量过程中,打开颈环结构,通过通用引物和特异引物,用SYBR染料,进行定量。 引物设计(仅供参考): 以>osa-miR171a为例: miR171a: UGAUUGAGCCGCGCCAAUAUC RT-primer: 5’GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGAgatattg3’ Forward primer: 5’gcgaTGATTGAGCCGCGCC3’ Reverse primer: 5’GTGCAGGGTCCGAGGT3’ 实验目的:检测水稻各种组织中的small RNA的表达量,特别是样品很少的材料。
关键词: Small RNA, Stem-loop RT, Real-time PCR, 颈环结构, SYBR
材料与试剂
仪器设备
实验步骤
参考文献
If you have any questions/comments about this protocol, you are highly recommended to post here. We will invite the authors of this protocol as well as some of its users to address your questions/comments. To make it easier for them to help you, you are encouraged to post your data including images for the troubleshooting.