Whole mount in situ localization of miRNAs and target mRNA transcripts in plants

被引:2
作者
Gautam, Vibhav [1 ,2 ]
Singh, Archita [1 ]
Verma, Swati [1 ]
Singh, Sharmila [1 ]
Chatterjee, Sourav [1 ]
Sarkar, Ananda K. [1 ]
机构
[1] Natl Inst Plant Genome Res, Aruna Asaf Ali Marg, New Delhi 110067, India
[2] Banaras Hindu Univ, Ctr Expt Med & Surg, Inst Med Sci, Varanasi 221005, Uttar Pradesh, India
关键词
In situ; miRNA; Gene expression; Arabidopsis; Root development; LOCKED NUCLEIC-ACID; HYBRIDIZATION; CELLS; GENE;
D O I
10.1007/s13205-019-1704-x
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Thefunctional characterization of miRNAs ofteninvolves understandingof their spatiotemporal expression, which mostly relies on reporter-based or in situ hybridization studies. The available in situlocalization methods follow separate protocols for pre-hybridization, hybridization, post-hybridization, and detection steps for both miRNA and mRNA transcripts in plants. In this study, we present a single method which can be used for whole mount in situ localization of both miRNAs and mRNAs in different plant tissues. Our modified method providesenhanced sensitivity for the localization of miRNA and their target transcripts. Consequently, a less laborious, time-saving, economic and efficient method has been proposed by the modification of pre-hybridization, hybridization, post-hybridization and detection steps.
引用
收藏
页数:7
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