Method for purifying viral nucleic acids
AssigneeNew York Blood Center, Inc.
InventorsDong-Hun Lee, Alfred M. Prince
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Abstract
The invention relates to a method for purification of viral RNA from a biological sample. The method involves lysing the virus envelope to liberate the RNA and passing the lysate through a porous hydrophilic PVDF filter to capture the viral RNA. The filter with bound RNA is then washed to remove proteins, lipids and other contaminants. The RNA is released from the filter using a low ionic strength ribonuclease (RNase) free solution to form a solution containing purified viral RNA. From this solution the RNA is recovered. The invention is also compatible with purification of nucleic acids from other types of samples.
Claims38
A method for purifying viral RNA, comprising:
- (a) applying a biological sample containing viral RNA to a hydrophilic polyvinylidine fluoride (PVDF) membrane which contains pores having an average diameter less than about 0.45 .mu.m;
- (b) passing the biological sample through the PVDF membrane;
- (c) washing the membrane to remove impurities while selectively retaining the viral RNA;
- (d) suspending the viral RNA using an eluant to provide purified viral RNA;
- and
- (e) recovering the purified viral RNA.
A method for purifying nucleic acid comprising:
- (a) applying a biological sample containing nucleic acid to a hydrophilic polyvinylidine fluoride (PVDF) membrane which contains pores having an average diameter less than about 0.45 .mu.m;
- (b) passing the biological sample through the PVDF membrane;
- (c) washing the membrane to remove impurities while selectively retaining the nucleic acid;
- (d) suspending the nucleic acid using an eluant to provide a purified sample;
- and
- (e) recovering the purified nucleic acid.
A method for purifying RNA relative to DNA in a sample comprising:
- (a) applying a sample containing RNA and DNA to a hydrophilic PVDF membrane which contains pore sizes having an average diameter of less than about 0.45 .mu.m;
- (b) passing the sample through the membrane to selectively retain the RNA;
- (c) washing the membrane;
- and
- (d) eluting to recover an eluate sample that has a substantially increased proportion of RNA to DNA.
Dependencies are inferred from the claim text.