DNA Affinity Chromatography Using Gravity Flow - Subscription Required DNA Affinity Chromatography, DNA affinity chromatography can be a low-tech method using gravity flow at 4Â°C, a disposable chromatography column, and DNA affinity resin prepared in the laboratory (see Preparation of a DNA Affinity Column). Include 10-20% glycerol and 0.025-0.1% NP-40 in the column buffers to suppress losses due to nonspecific adsorption of protein to surfaces. Load the protein in a buffer that is compatible with binding of the protein to its target site. Keith Brocklehurst et al
Selection of Poly(A)+ RNA by Batch Chromatography - Subscription Required When many RNA samples are to be processed or when working with small amounts (<50 Âµg) of total mammalian RNA, the technique of choice is batch chromatography on oligo(dT)-cellulose. The method described in this protocol uses a combination of temperature and ionic strength to maximize binding and recovery of polyadenylated RNA. IMPORTANT: Prepare all reagents used in this protocol with Diethyl pyrocarbonate (DEPC)-treated H2O. Joseph Sambrook and David W. Russell.
Selection of Poly(A)+ RNA by Oligo(dT)-Cellulose Chromatography - Subscription Required Chromatography on oligo(dT) columns is the preferred method for large-scale purification (>25 Âµg) of poly(A)+ RNA extracted from mammalian cells. Typically, between 1% and 10% of the RNA applied to the oligo(dT) column is recovered as poly(A)+ RNA. Because the method can be frustratingly slow, it is not recommended for purification of poly(A)+ RNA from multiple samples. For this purpose, batch elution (Selection of Poly(A)+ RNA by Batch Chromatography) is the better choice.
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