Intended use
In this kit, guanidine isothiocyanate is used for pyrolysis, total DNA and RNA in samples are separated and purified by magnetic beads adsorption, and PCR inhibitors such as proteins, lipids and ribonuclease can be effectively removed after purification. The purified products can be directly used for fluorescence quantitative PCR.
Inspecting principle
In this kit, guanidine isothiocyanate is used for lysis, nucleic acid is released, nucleic acid is absorbed by electrostatic effect, hydrophobic effect and hydrogen bond effect, and PCR inhibitors such as proteins, lipids and ribonuclease are removed after washing.
Main components
Reagent prepackaged 96 well plate
Storage conditions and period of validity
The kit was stored at 4℃, Carrier RNA were stored at -20℃, and the expiry date was 12 months. Heat preservation packaging shall be used for transportation, and pre-cooled low-temperature ice bags shall be added at -20℃, with the amount of ice bags not less than 20% of the total volume. Sealed container shall be transported for no more than 72 hours.
Product Performance Index
According to clinical assessment, the DNA detection sensitivity of this kit is 500IU/mL;The detection sensitivity of RNA was 500IU/mL.Hemolysis, hyperlipidemia, high bilirubin and high immunoglobulin samples should be avoided in this kit.
Extraction Method
1.Preparation of reagent board
(1)Take out the prepackaged reagent board from the kit and turn it upside down several times to make the magnetic beads suspend evenly;
(2)Carefully tear off the aluminum foil sealing film to avoid the vibration of orifice plate and prevent the liquid from splashing out.
2.Sample preparation
100 μ l of sample was added to deep well plate station 1, and 10 μ l of protease K and 2 μ l of carrier RNA were added to the cleavage tank.
3.On machine extraction
put the prepared 96 deep well plate in the nucleic acid extractor, insert the magnetic rod sleeve, and extract.
| Step | Slot position | Name | Magnetic absorption time | Mixing speed | Time | Temperature |
| 1 | 2/8 | Magnetic beads | 0s | low | 0s | off |
| 2 | 3/9 | lysis | 60s | low | 600s | 60℃ |
| 3 | 4/10 | wash | 30s | fast | 0s | off |
| 4 | 6/12 | elute | 30s | fast | 0s | off |
Precautions
1.Before the experiment, the kit instructions should be read in detail.
2.In the process of specimen processing, the supernatant removal steps involved, please try to absorb the supernatant, while avoiding to absorb the precipitate, so as not to affect the accuracy of detection.
3.Samples should be considered as potential sources of infection, and the operation and treatment should meet the requirements of relevant regulations: General Guidelines for Biosafety of Microbial and Biomedical Laboratories and Regulations on the Management of Medical Wastes issued by the Ministry of Health.
