Agarose gel electrophoresis of bacteriophages and related particles. I. Avoidance of binding to the gel and recognizing of particles with packaged DNA
Agarose gel electrophoresis of bacteriophages and related particles. I. Avoidance of binding to the gel and recognizing of particles with packaged DNA
复制标题
噬菌体和相关颗粒的琼脂糖凝胶电泳。
作者：
P. Serwer;S. Hayes
使用琼脂糖凝胶电泳法分离病毒和相关颗粒的两个潜在障碍是：(A)在电泳过程中样品与凝胶的粘附性，以及(B)确定被核酸特异染色的颗粒是否具有包装状态的核酸所需的冗长程序。已经证明，噬菌体T7的带正电的尾部纤维附着在琼脂糖凝胶上是前面描述的(Serwer，P.和Pichler，M.E.，J.Virol)的可能原因。1978，28,917-928)在琼脂糖凝胶电泳法(浓度效应)过程中，噬菌体T7形成的条带的扩散依赖于样品浓度。描述了用于筛选琼脂糖制剂以防止颗粒粘连的方法，并且使用这些方法已经发现，在设计用于低电渗透的琼脂糖制剂中，T7的浓度效应被减小或消除。此外，还表明，在电泳法后，T7噬菌体(和其他DNA噬菌体)对核酸特异的菌株溴化乙锭的染色强度因从T7内部喷射DNA而增加；这种染色强度的增加用于识别包装DNA的颗粒形成的条带。
Two potentialobstacles to using agarose gel electrophoresis for the fractionation of viruses and related particles are: (a) adherence of the sample to the gel during electrophoresis, and (b) the prolonged procedures necessary for determining whether or not particles stained with nucleic acid‐specific stains have their nucleic acid in a packaged state. It is demonstrated that adherence of positively charged tail fibers of bacteriophage T7 to agarose gels is the probable cause of a previously described (Serwer, P. and Pichler, M. E., J. Virol. 1978, 28, 917–928) sample concentration‐dependent spreading of bands formed by bacteriophage T7 during agarose gel electrophoresis (concentration effect). Procedures for screening preparations of agarose for nonadherence of particles are described and use of these procedures has resulted in the finding that the concentration effect is decreased or eliminated for T7 in preparations of agarose designed for low electro‐osmosis. In addition, it has been shown that after electrophoresis, the intensity with which bacteriophage T7 (and other DNA bacteriophages) stains with the nucleic acid‐specific stain, ethidium bromide, is increased by ejecting DNA from the interior of T7; this increase in staining intensity is used to recognized bands formed by particles with packaged DNA.
影响因子：
2.9
作者：
Serwer,P
通讯作者：
Serwer,P