Protocol from:(Zhizhong Gong et al. 2005)(Parry et al. 2006)
Oligo d(T) should be 45-50nt long and labeled in 5’. Do not use fluorophore in the red range has they would be mask by chlorophyll. Alexa-488 is a good choice (Invitrogen).
120 mM | NaCl |
7 mM | Na2HPO4 |
3 mM | NaH2PO4 |
2.7 mM | KCl |
0.1% | Tween 20 |
80 mM | EGTA |
5% | formaldehyde |
10% | DMSO |
DAY1
Approx 4h
Wash twice times 5 minutes in ethanol
Gong, Zhizhong, Chun-Hai Dong, Hojoung Lee, Jianhua Zhu, Liming Xiong, Deming Gong, Becky Stevenson, and Jian-Kang Zhu. 2005. A DEAD box RNA helicase is essential for mRNA export and important for development and stress responses in Arabidopsis. The Plant Cell 17, no. 1 (January): 256-67. doi:10.1105/tpc.104.027557.
Parry, Geraint, Sally Ward, Alex Cernac, Sunethra Dharmasiri, and Mark Estelle. 2006. The Arabidopsis SUPPRESSOR OF AUXIN RESISTANCE proteins are nucleoporins with an important role in hormone signaling and development. The Plant Cell 18, no. 7 (July): 1590-603. doi:10.1105/tpc.106.041566.