RNA Isolation from High-Phenolic Tea Leaves and Apical Buds

被引:18
作者
Lal L. [1 ]
Sahoo R. [1 ]
Gupta R.K. [1 ]
Sharma P. [2 ]
Kumar S. [1 ]
机构
[1] Biotechnology Division, Inst. of Himalyan Biorsrc. Technol., Palampur-176 061 (HP)
[2] Div. of Agricultural Biotechnology, Himachal Pradesh Krishi V.
关键词
Camellia sinensis; Caragana jubata; Phenolics; Potentilla astrisanguinea; RNA extraction;
D O I
10.1007/BF02772161
中图分类号
学科分类号
摘要
We describe here a modified procedure based on guanidine hydrochloride to isolate RNA from apical buds of tea containing high levels of polyphenols. The major change was to increase β-mercaptoethanol from 50 to 200 mM in the extraction buffer. This produced phenolic-free and undegraded RNA. The procedure was extended to fully expanded leaves of tea and apical buds of other plant species. The total RNA could be used for mRNA isolation, reverse transcription, and differential display of mRNA. A clean-up protocol to remove chlorophylls that coprecipitate with the RNA also is presented.
引用
收藏
页码:181a / 181f
相关论文
共 9 条
[1]  
Dawson R.M.C., Elliott D.C., Elliott W.H., Jones K.M., Data for Biochemical Research, pp. 380-381, (1986)
[2]  
Liang P., Zhu W., Zhang X., Guo Z., O'Connell R.P., Averboukh L., Wang F., Pardee A.B., Differential display using one-base anchored oligo-dT primers, Nucleic Acids Res, 22, pp. 5763-5764, (1994)
[3]  
Logemann J., Schell J., Willmitzer L., Improved method for the isolation of RNA from plant tissues, Anal Biochem, 163, pp. 16-20, (1987)
[4]  
Loomis W.D., Overcoming problems of phenolics and quinones in the isolation of plant enzymes and organelles, Meth Enzymol, 31, pp. 528-545, (1974)
[5]  
Manning K., Isolation of nucleic acids from plants by differential solvent precipitation, Anal Biochem, 195, pp. 45-50, (1991)
[6]  
Salzman R.A., Fujita T., Zhu-Salzman K., Hasegawa P.M., Bressan R.A., An improved RNA isolation method for plant tissues containing high levels of phenolic compounds or carbohydrates, Plant Mol Biol Reptr, 17, pp. 11-17, (1999)
[7]  
Sambrook J., Fritsch E.F., Maniatis T., Molecular Cloning: A Laboratory Manual, (1989)
[8]  
Venkatachalam P., Thanseem I., Thulaseedharan A., A rapid and efficient method for isolation of RNA from bark tissues of Hevea brasiliensis, Curr Sci, 77, pp. 635-637, (1999)
[9]  
Yamanishi T., Chemical composition of tea, Food Rev Int, 11, 3, pp. 435-456, (1995)