A method for isolating nucleic acid which comprises:
- (a) applying a sample comprising cells containing nucleic acid to a
filter, whereby the cells are retained as a retentate and contaminants are removed;
- (b) lysing the retentate from step (a) while the retentate is retained
by the filter to form a cell lysate containing the nucleic acid;
- (c) filtering the cell lysate with the filter to retain the nucleic
acid and remove remaining cell lysate;
- (d) optionally washing the nucleic acid retained by the filter; and
- (e) eluting the nucleic acid, wherein the filter composition and dimensions
are selected so that the filter is capable of retaining the cells and the nucleic acid.
Additionally, there is provided a substrate for lysing cells and purifying
nucleic acid having a matrix and a coating and an integrity maintainer for maintaining
the purified nucleic acid. Also provided is a method of purifying nucleic acid
by applying a nucleic acid sample to a substrate having an anionic detergent affixed
to a matrix, the substrate physically capturing the nucleic acid, bonding the nucleic
acid to a substrate and generating a signal when the nucleic acid bonds to the
substrate indicating the presence of the nucleic acid. A kit for purifying nucleic
acid containing a coated matrix and an integrity maintenance provider for preserving
the matrix and purifying nucleic acid is also provided. Further, there is provided
a method for quantifying DNA, such as double-stranded or genomic DNA, isolated
from cells, such as leukocytes to determine the numbers of leukocytes in a sample
of leukoreduced blood.
|
|
|