Dot and Slot Hybridization of Purified RNA Protocol Protocol for dot and slot hybridization of purified RNA. Dot blotting of RNA is best carried out using purified preparations of RNA that are denatured with glyoxal or formaldehyde immediately before loading onto a nylon membrane through a vacuum manifold.
Northern Blot Analysis of mRNA From Mammalian Polyribosomes Protocol The protocol consists of a method for the generation of cytoplasmic extracts from mammalian cells (in this case, 293T cells) without the disruption of polyribosomes, the separation of ribosomal components and polyribosomes by sucrose gradient centrifugation, the isolation of mRNA from these fractions, and detection of mRNA by Northern blot analysis.
Northern Hybridization Protocol Protocol for northern hybridization. Protocol describes how to carry out northern hybridization at high stringency in phosphate-SDS-buffers. Although a wide variety of formats are available, hybridization is usually performed in heat-sealable bags, roller bottles, or plastic boxes, as described here.
Southern Blotting: Capillary Transfer of DNA to Membranes Protocol Protocol describes the first stages of Southern blotting: digestion of genomic DNA with one or more restriction enzymes, separation of the resulting fragments by electrophoresis through an agarose gel, and transfer of the denatured fragments to a membrane by downward capillary transfer.
Southern Blotting: Simultaneous Transfer of DNA from a Single Agarose Gel to Two Membranes Protocol Protocol for southern blotting: simultaneous transfer of DNA from a single agarose gel to two membranes. DNA can be simultaneously transferred from opposite sides of a single agarose gel to two membranes. Bidirectional transfer occurs rapidly at first, but soon slows down as the gel becomes dehydrated. Because the efficiency of transfer is low, the method works best when the target sequences are present in high concentration
Southern Hybridization of Radiolabeled Probes to Nucleic Acids Immobilized on Membranes Protocol Protocol for southern hybridization of radiolabeled probes to nucleic acids immobilized on membranes. Protocol describes how to carry out Southern hybridizations at high stringency in phosphate-SDS buffers. Although a wide variety of formats are available, most Southern hybridizations are carried out in heat-sealable bags, roller bottles, or plastic boxes.
