Optimized Hybridization of DNA Blotted and Fixed to Nitrocellulose and Nylon Membranes

Seven nylon membranes were tested in different DNA transfer and post–transfer procedures. The sensitivity of the hybridization signals was increased ten–fold over conventional Southern blot analysis using nitrocellulose, by UV–irradiating DNA blotted to nylon membranes at high salt concentrations. This treatment also improved the sensitivity of the signals with nitrocellulose membranes. UV–irradiation of DNA transferred to nylon membranes in low salt buffers or alkali reduced hybridization efficiency

Protein Components of the microRNA Pathway and Human Diseases

MicroRNAs (miRNAs) are key regulators of messenger RNA (mRNA) translation known to be involved in a wide variety of cellular processes. In fact, their individual importance is reflected in the diseases that may arise upon the loss, mutation or dysfunction of specific miRNAs. It has been appreciated only recently that diseases may also develop when the protein components of the miRNA machinery itself are affected. The core enzymes of the major protein complexes involved in miRNA biogenesis and function, such as the ribonucleases III (RNases III) Drosha and Dicer as well as Argonaute 2 (Ago2), appear to be essential. However, the accessory proteins of the miRNA pathway, such as the DiGeorge syndrome critical region gene 8 (DGCR8) protein, Exportin-5 (Exp-5), TAR RNA binding protein (TRBP) and fragile X mental retardation protein (FMRP), are each related, in various ways, to specific genetic diseases