Solid medium and process for the storage and rapid purification of nucleic acid
a technology applied in the field of solid medium and process for the storage and rapid purification of nucleic acid, can solve the problems of biomolecule denaturation and deactivation, inability to retrieve covalently bound molecules from the filter membrane, and slow molecule immobilization,
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example 1
Heat Elution
78. Several drops of freshly finger-stick drawn blood was spotted to the filter membrane of the invention and allowed to air-dry for two minutes. Once dried two 1 mm diameter punches were taken from the dried blood spot and applied to individual 2000 ul polypropylene PCR tubes. To each tube containing a single 1 mm blood punch, 200 ul of FTA Purification Reagent (Fitzco, Inc) was added. Per 500 ml: 0.29 g NaCl; 5 ml 1 M Tris pH 7.5; 1 ml 0.5 M EDTA; 2.5 ml Triton x-100. Tubes were incubated for five minutes at room temperature with no shaking. Following incubation the FTA purification Reagent was aspirated from the tube. A second aliquot of 200 ul of FTA Purification Reagent was added to each tube. The tubes were incubated for five minutes at room temperature without shaking. Following incubation the FTA Purification Reagent was aspirated from both tubes. 200 ul of TE (10 mM Tris-HCl, 1 mM EDTA, pH 8.0) buffer was then added to each tube. The tubes were incubated for fiv...
example 2
Full Elution Protocol
80. Several drops of freshly finger-stick drawn blood were spotted to the filter membrane of the invention and allowed to air-dry for two minutes. Once dried two 1 mm diameter punches were taken from the dried blood spot and applied to individual 200 ul polypropylene PCR tubes. To each tube containing a single 1 mm blood punch, 200 ul of FTA Purification Reagent (Fitzco, Inc) was added. Tubes were incubated for five minutes at room temperature with no shaking. Following incubation the FTA purification Reagent was aspirated from the tube, 20 ul of the aspirate was retained. A second aliquot of 200 ul of FTA Purification Reagent was added to each tube. The tubes were incubated for five minutes at room temperature without shaking. Following incubation the FTA Purification Reagent was aspirated from both tubes, 20 ul of the aspirate was retained. 200 ul of TE buffer was then added to each tube. The tubes were incubated for five minutes at room temperature without sh...
example 3
Comparison of Elution
83. Single stranded DNA can be readily detected with the use of OliGreen.RTM. (Molecular Probes, Inc), a fluorescent probe specific for the single stranded molecule. By using OliGreen the total single stranded DNA eluted from the filter membrane of the invention can be determined, as well as other single stranded DNA purification methods. A standard curve for single stranded genomic DNA was constructed according to manufacture's instructions (Molecular Probes, Inc) (see FIG. 3a).
84. 5 ul of freshly finger-stick drawn blood was spotted to a 7 mm disk of the filter membrane, composed of a chemically coated porous glass microfiber filter membrane, of the invention and also to a 7 mm disk of the commercially available FTA solid support. Both spots were allowed to air-dry for two minutes. Once dried, the punches were applied to individual 1.5 ml polypropylene Eppendorf tubes. To each tube containing a single 7 mm blood punch, 1 ml of FTA Purification Reagent (Fitzco,...
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Abstract
Description
Claims
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