A simple and rapid method for transferring RNA from agarose gels to nitrocellulose paper for blot hybridization has been developed. Poly(A)+ and ribosomal RNAs transfer efficiently to nitrocellulose paper in high salt (3 M NaCI/0.3 M trisodium citrate) after denaturation with glyoxal and 50% (vol/vol) dimethyl sulfoxide. RNA also binds to nitrocellulose after treatment with methylmercuric hydroxide. The method is sensitive: about 50 pg of specific mRNA per band is readily detectable after hybridization with high specific activity probes (108 cpm/sg). The RNA is stably bound to the nitrocellulose paper by this procedure, allowing removal of the hybridized probes and rehybridization of the RNA blots without loss of sensitivity. The use of nitrocellulose paper for the analysis of RNA by blot hybridization has several advantages over the use of activated paper (diazobenzyloxymethyl-paper). The method is simple, inexpensive, reproducible, and sensitive. In addition, denaturation of DNA with glyoxal and dimethyl sulfoxide promotes transfer and retention of small DNAs (100 nucleotides and larger) to nitrocellulose paper. A related method is also described for dotting RNA and DNA directly onto nitrocellulose paper treated with a high concentration of salt; under these conditions denatured DNA of less than 200 nucleotides is retained and hybridizes efficiently. The technique of Southern (1) enables one to transfer electrophoretically separated DNA fragments to nitrocellulose paper for hybridization with specific radioactive DNA or RNA probes. Although RNA does not generally bind to nitrocellulose, it has been transferred and covalently coupled to activated cellulose paper (diazobenzyloxymethyl-paper, DBM-paper) according to the method of Alwine et al. (2,3). In our laboratory, the use of activated paper for coupling RNA has been complicated by two major problems: first, about 500 pg of specific RNA per band is just detectable (after several days exposure) after hybridization using high specific activity probes prepared by nick translation (108 cpm/,ug). Although this sensitivity is similar to that obtained by others with this method (2, 3), we estimate that this is 1-10% of that for detecting specific DNA sequences on nitrocellulose by using similar probes. The increased sensitivity of the nitrocellulose paper is probably a reflection of the fact that nitrocellulose has a higher binding capacity for DNA (about 80 ,Ug/cm2) compared to the capacity of most preparations of activated paper for binding ,gg/cm2). Second, we have found that preparation and activation of DBM-paper is expensive, time consuming, and, most importantly, often variable.Poly(A)+ RNA is retained on Millipore filters in 0.5 M KCI (4, 5). We therefore decided to investigate whether RNA in general could be bound to nitrocellulose paper and retained during hybridization and stringent washing. Here I describe a rapid and simple method for blot hybridization of denatured The publication costs of this article were defrayed in part by page charge pa...

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The purpose of this article is to describe a preadmission, preoperative educational program offered free of charge for patients undergoing total joint replacement surgery at a large teaching hospital located in metropolitan Detroit, Michigan. In establishing the preoperative educational program, a multidisciplinary approach was used to provide a comprehensive learning environment for patients and their families. To evaluate the effectiveness of the program, patients completed surveys at the end of each class. Patients reported that their expectations of the program were met, they were less anxious about their surgery as a result of attending the classes, and the preoperative teaching by the multidisciplinary team was effective. Having a live session that offered an opportunity to ask individual and specific questions to each healthcare professional with immediate feedback proved to be a positive experience for patients. Patients' comments supported the multidisciplinary team's impression that real-time, interactive teaching was highly valued by patients and their caregivers.