Nucleic acid extraction from complex matrices
The present disclosure describes an adsorbent and exemplary protocols for extracting nucleic acids, such as DNA and RNA, from complex matrices, such as stool samples and water samples. The adsorbent is activated charcoal coated with a material such as polyvinylpyrrolidone, dextran, or coconut flours. The adsorbent may be used in microcentrifuge spin columns, where it may be present as a slurry in a storage solution. The sample may be prepared by vortexing in a buffer solution, centrifuging, adding a protease to the supernatant, and passing the supernatant through a microcentrifuge spin column containing coated activated charcoal. The key components, including buffer, protease, and spin columns, may be packaged in a kit.
1. A process for extracting nucleic acids from a sample, the process comprising: homogenizing at least a portion of the sample in a buffer; centrifuging the homogenized sample portion, the centrifugation resulting in a pellet and a supernatant; separating the supernatant from the pellet; adding a protease enzyme to the supernatant; and passing the supernatant over an adsorbent, the adsorbent comprising a slurry of about 5% to about 20% weight/volume activated charcoal in a storage solution, the activated charcoal being coated with a coating material selected from the group consisting of polyvinylpyrrolidone, dextran, and coconut flours.
2. The process of claim 1 , wherein the sample is selected from the group consisting of a stool sample, a tissue sample, a urine sample, a blood sample, a water sample, a soil sample, an agricultural sample, a veterinary sample, and a food sample.
3. The process of claim 1 , wherein the portion of the sample has at least one of a mass of 0.2 g or a volume of 0.2 μl.
4. The process of claim 1 , further comprising incubating the supernatant.
5. The process of claim 1 , wherein the slurry comprises from about 1% to about 10% weight/volume of the coating material.
6. A kit for extracting nucleic acids from a sample the kit comprising:
a container of buffer solution;
a container of protease enzymes; and
at least one spin column comprising a slurry of about 5% to about 20% weight/volume coated activated charcoal, wherein the coating material is about 1% to about 10% weight/volume polyvinylpyrrolidone.
7. The kit of claim 6 , wherein the buffer solution is configured to homogenize the sample when a portion of the sample is placed in the buffer and vortexed.
8. The kit of claim 6 , wherein the coated activated charcoal is configured to bind to contaminant while leaving a nucleic acid unbound.
9. An adsorbent comprising a slurry of about 5% to about 20% weight/volume activated charcoal in a storage solution, the activated charcoal being coated with a coating material selected from the group consisting of polyvinylpyrrolidone, dextran, and coconut flours.
10. A microcentrifuge column comprising the absorbent of claim 9 .