Transfusion medicine leukodepletion filter devices as a source of genetic material for genotyping studies
AssigneeWhatman, Inc.
InventorsMartin A. Smith, Galina N. Fomovskaia, Mikhail A. Fomovsky, Neil J. Butt, Matthew Baker
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Abstract
The present invention provide methods for utilizing spent leukodepletion filter devices as a source of material for the isolation and analysis of genomic DNA (gDNA), including polymorphism, genotyping, and pharmacogenomic studies. Cellular retentate with the filter contains leukocytes, which are lysed to release the nuclei. The nuclei are lysed or ruptured to release genomic DNA, which is then isolated and used for subsequent analysis.
Claims34
A method for isolating DNA comprising:
- (a) providing a leukodepletion filter device having cellular retentate containing leukocytes;
- (b) lysing the cellular retentate to form a cell lysate;
- (c) treating the filter to remove cell lysate from the filter while retaining leukocyte nuclei with the filter;
- (d) rupturing leukocyte nuclei retained with the filter in step (c) and removing contaminants while retaining the nucleic acid;
- and
- (e) eluting the nucleic acid from the filter.
A method for utilizing leukodepletion filter devices as a source for the isolation of nucleic acid, the method comprising:
- (a) providing a leukodepletion filter device having cellular retentate containing leukocytes containing nucleic acid;
- (b) lysing the cellular retentate to form a cell lysate;
- (c) treating the filter to remove cell lysate from the filter while retaining leukocyte nuclei with the filter;
- (d) rupturing leukocyte nuclei retained with the filter in step (c) and removing contaminants while retaining the nucleic acid;
- (e) heating the filter retaining the nucleic acid to an elevated temperature of 40° C. to 125° C.;
- and
- (f) eluting the nucleic acid.
Dependencies are inferred from the claim text.