Method for recovering sperm nucleic acid from a forensic sample
A method for selectively recovering nucleic acid from a sperm cell in a sample containing cells of at least a sperm cell and an epithelial cell, and a cell suspension medium comprising extracellular impurities, is provided. The method entails introducing a sample into a vessel, sequestering the cells from the remaining sample components, washing the cells with a washing solution either before or after sequestration, removing the impurities-containing cell suspension medium from the vessel while retaining the cells; lysing selectively cells of the first cell type; and isolating the nucleic acid from the lysed cells. Methods for recovering nucleic acid from the second cell type are also provided.
1. A method for selectively recovering DNA from sperm cells from a sample containing intact sperm cells and intact epithelial cells, the method comprising the steps of:
(a) washing a sample comprising intact sperm cells and intact epithelial cells by mixing the sample with a washing solution, wherein the washing solution comprises from 30 mM to 260 mM MgCl 2 and thereby forming a washed sample;
(b) applying DNase to the washed sample and thereby forming a DNased sample;
(c) applying a selective sperm lysis buffer comprising NaCl and DTT to the DNased sample of step (b); and
(d) isolating sperm DNA from the sample of step (c).
2. The method of claim 1 , wherein 0.01 U to 50 U of DNase per 100 uL is applied to the washed sample.
3. The method of claim 1 , comprising purifying the isolated sperm DNA to a purity sufficient for sperm short tandem repeat (STR) profiles to be obtained that are suitable for genotyping.
4. The method of claim 1 , wherein the washing solution comprises from 160 mM to 260 mM MgCl 2 .