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Surgical Scalpel Blade No.18


Company: Swann Morton
Catalog#: 0323
Other protocol()

An Efficient Procedure for Protoplast Isolation from Mesophyll Cells and Nuclear Fractionation in Rice
[Abstract]  Plant protoplasts, a proven physiological and versatile cell system, are widely used in high-throughput analysis and functional characterization of genes. Green protoplasts have been successfully used in investigations of plant signal transduction pathways related to hormones, metabolites and environmental challenges. This protocol, adapted from Zhang et al. (2011), describes a procedure for the isolation of rice protoplasts from green tissue and shows an efficient and rapid method for isolation of nuclei form these protoplasts which are commonly used in a variety of experimental procedures including the isolation of high-molecular-weight DNA (Watson and Thompson, 1986), in vitro DNA synthesis (Roman, 1980), isolation of labeled transcripts for differential screening of ... [摘要]  植物原生质体,经证实的生理和多功能细胞系统,广泛用于基因的高通量分析和功能表征。绿色原生质体已经成功地用于调查与激素,代谢物和环境挑战相关的植物信号转导途径。该协议改编自Zhang等人(2011),描述了从绿色组织中分离水稻原生质体的方法,并且显示了用于从这些原生质体中分离核的有效和快速的方法,所述原生质体是常用的在包括分离高分子量DNA(Watson和Thompson,1986),体外 DNA合成(Roman,1980)的各种实验过程中,分离用于cDNA的差异筛选的标记的转录物图书馆(Somssich等人,1989),用于体外转录系统的核提取物的制备(Roberts和Okita,1991),核蛋白的分离(Harrison& et al。,1992)和蛋白质靶向核的研究(Hicks和Raikhel,1993)。