◎ The operation is simple and rapid, and the extraction of microbial nucleic acid in the sample can be completed in about 20 minutes;
◎ The extracted nucleic acid has high purity and no inhibitor, 1.75.
◎ The unique lysate formula is suitable for the extraction of microbial nucleic acid from blood, saliva, swab, water sample and feces, especially for the molecular detection of microbial infection in livestock.
Product introduction
BIOG microbial nucleic acid extraction kit is repeatedly developed and optimized by R&D team of our company on the basis of comprehensive comparison with similar foreign high-quality products. It is specially used for rapid extraction and purification of microbial nucleic acids in samples, especially for molecular detection of microbial infections in domestic animals such as pigs, cattle and sheep. The kit adopts a unique lysate formula, which can directly extract high-quality microbial nucleic acid from fresh, frozen or old samples such as whole blood, swab, saliva, water sample, feces and soil. The operation is simple and fast, and the extraction of microbial nucleic acid can be completed in about 20 minutes. The extraction rate of nucleic acid is high, and about 2-4μg of nucleic acid can be extracted from 200μL whole blood. BIOG microbial nucleic acid extraction kit adopts the latest high-quality ionic membrane, and the lysate and eluent have been optimized for many times to remove impurities such as protein, pigment and lipid to the maximum extent, which is especially suitable for molecular detection of microbial infections including bacteria and viruses in samples.
Operating steps? (Please prepare anhydrous ethanol, isopropanol and sterilized 1.5mL centrifuge tubes by yourself):
1. Take out the washing liquid and do the following:
A) Wash solution A: 2 1 ml, and add 9mL of absolute ethanol; Add 18mL absolute ethanol into 42mL and mix well.
B) Wash solution B: 18 ml, and add 42 ml of anhydrous ethanol; Add 84 ml of anhydrous ethanol to 36 ml and mix thoroughly.
C) If the prepared washing liquid has precipitation, it can be dissolved at 37℃ and shaken before use.
2. Take a 1.5mL centrifuge tube, add 200μL of blood or other samples, then add 200μL of lysate and 40μL of digestive juice, shake well, and take a water bath at 65℃10min.
3. Add 150μL isopropanol (25% of the total volume) and mix well. If there is translucent suspended matter, it will not affect DNA extraction and subsequent experiments.
4. Put the adsorption column into the collection tube, transfer the above solution into the adsorption column, centrifuge at 12000 rpm 1 min, and discard the waste liquid in the collection tube;
5. Put the adsorption column back into the collection tube, add 500μL of washing liquid A to the adsorption column, let it stand for 65438 0 minutes, centrifuge at 65438±02000 rpm for 65438 0 minutes, and discard the waste liquid in the collection tube.
6. Put the adsorption column back into the collection tube, add 500μL of washing liquid B to the adsorption column, centrifuge at 65438±02000 rpm for 65438 0 minutes, and discard the waste liquid in the collection tube.
7. Put the adsorption column back into the collection tube, add 500μL of washing liquid B to the adsorption column, centrifuge at 65438±02000 rpm for 65438 0 minutes, and discard the waste liquid in the collection tube.
8. Take 30μL eluent from each sample (for example, take 300μL eluent from 10 sample), put it in a sterilized 1.5mL centrifuge tube, and preheat it at 65℃.
9. Put the adsorption column back into the collection tube and centrifuge at 12000 rpm for 2 minutes, leaving residual washing liquid.
10. Take out the adsorption column, put it into a new 1.5 mL sterile centrifuge tube, add 30 μL preheated eluent to the center of the adsorption column, let it stand for 2min, centrifuge at 12000 rpm 1min, and collect nucleic acid solution. The extracted nucleic acid can be used in the next experiment or stored at -20℃.
Precautions:
1. The dissolved product contains irritating chemicals. Please take protective measures during operation to avoid direct skin contact and inhalation. If the cracking liquid is not used up after unpacking, tighten the pipe in time.
Cover it and keep it sealed.
2. The dosage of a single blood sample is generally 200μL l L. If there is a large amount of blood, please use the BIOG medium whole blood nucleic acid extraction kit.
3. In order to prevent RNase pollution, it is best to wear disposable cleaning gloves and masks, and use treated containers and ultra-pure water without RNase.
4. If the extracted DNA is not used for the time being, it should be stored at -20℃, RNA should be stored at -70℃, and RNA protection factors can be appropriately added (article number: 5 1090, generally 50μ L LRNA solution plus 2μL).