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US 9,809,794 B2Granted
Materials and methods for achieving differential lysis of mixtures with the aid of alkaline lysis and pressure cycling technology (PCT)
AssigneesThe Florida International University Board of Trustees, Pressure Biosciences, Inc.
InventorsBruce R. McCord, Deepthi V. Nori, Nathan P. Lawrence, Richard Schumacher
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Abstract
The subject invention provides a two-step protocol using pressure cycling technology (PCT) and alkaline lysis for differential extraction of mixtures. In a preferred embodiment the procedure is used in forensic DNA applications such as, for example, DNA testing in the case of rape.
Claims14
A method of separating male sperm cells and female epithelial cells in a sample embedded in a cotton swab, comprising:
- immersing the sample in an alkaline solution comprising 0.2-0.8 N NaOH,
- wherein said alkaline solution does not comprise deoxyribonuclease (DNase);
- applying pressure cycling technology (PCT) to the sample at room temperature, the PCT comprising the application of alternating cycles of ambient and high pressure to the sample, said high pressure being between 10,000 psi and 45,000 psi;
- neutralizing the sample with a buffer solution;
- centrifuging the sample to produce an eluted epithelial fraction;
- collecting and purifying the eluted epithelial fraction for female DNA processing, whereby a sperm fraction is left in the cotton swab;
- and
- processing the sperm fraction remaining in the cotton swab.
5 more elements
Dependencies are inferred from the claim text.