A rapid staining technique for detection of RNase after polyacrylamide gel electrophoresis

✍ Scribed by Curtis W. Wilson

• Publisher: Elsevier Science
• Year: 1969
• Tongue: English
• Weight: 363 KB
• Volume: 31
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(69)90294-2

✦ Synopsis

Methods for the detection of RNase after polyacrylamide gel electrophoresis have required long incubation times or indirect techniques that allowed the enzyme ample time to diffuse into a broad band (l-3). Wolf (4) recently described a new technique in which the gel was incubated in a solution of low molecular weight RNA, which diffused into the gel. The gel was then stained with 0.1% methylene blue." This method produced sharp bands at the site of RNase activity, but required 2 hours of incubation, 2 hours of staining, and at least 12 hours for destaining.

Randles (5) cast the gel with high molecular weight RNA, inhibited RNase during electrophoresis, removed the inhibitor, then incubated to allow the enzyme to act on the RNA. This was followed by fixing, staining with toluidine blue, and destaining.

I wish to report a procedure for the detection of RNase in polyacrylamide gels which requires much less time than the methods reported above, and which is sensitive to small amounts of plant RNase.