NUCLEIC ACID PURIFICATION

摘要

Methods and composition for nucleic acid isolation are provided. In one embodiment, a method is provided for nucleic acid purification from biological samples, such as whole blood samples, extracted with phenol-based denaturing solvents, which does not require phase separation or nucleic acid precipitation. Methods according to the invention may also be used for of small RNAs (e.g., siRNAs or miRNAs) purification and are amenable to automation.

主权项

1. A method for binding a nucleic acid to a silica substrate for purification comprising:
(a) contacting sample comprising whole blood or blood cells with a denaturing solvent comprising phenol; (b) adding a binding agent to the sample, wherein the binding agent comprises a chaotropic salt, a lower alcohol or a mixture thereof; and (c) contacting the sample with a silica substrate thereby binding the nucleic acid to the silica substrate,wherein the nucleic acid in the sample is substantially unprecipitated prior to binding of the nucleic acid to the silica substrate, andwherein the nucleic acid is bound to the silica substrate with out substantially separating the organic and aqueous phases in the sample.