Blood DNA Kit Oral swab DNA extraction kit (centrifugal column type)

User Manual

Product Name

Common name: Blood DNA Kit Oral Swab DNA extraction Kit (centrifugal column type)

Trade name: Blood DNA Kit Oral swab DNA extraction kit (centrifugal column type)
  

Intended use

This kit uses a centrifugal adsorption column that can specifically bind DNA and a unique buffer system to extract DNA from oral swab samples. The silicon matrix material used in the centrifugal adsorption column can efficiently and specifically adsorb DNA, and combined with a unique buffer system, it can maximize the removal of impurity proteins and other organic compounds in the cell. The extracted genomic DNA fragments are complete, highly purified, and of stable and reliable quality. The purified DNA using this kit is suitable for various conventional experiments such as enzyme digestion, PCR, library construction, Southern hybridization, and experiments that require high-quality DNA such as chip hybridization and high-throughput sequencing.
  

Inspection principle

1. For DNA extraction of oral swabs, the samples can be fresh or frozen.

2. High quality: Unique lysis buffer system, purified DNA with high concentration, high purity, and good integrity, meeting experimental requirements such as chip hybridization and high-throughput sequencing.

3. Fast and non-toxic: Adopting the principle of silica gel membrane adsorption, the experiment can be completed within 1 hour without the need for phenol chloroform.

Precautions (please be sure to read this note before using this reagent kit):

1. The sample should avoid repeated freeze-thaw, otherwise it may result in a smaller extracted DNA fragment and a decrease in extraction volume. Extracting samples requires balancing to room temperature.

If there is sediment in the cracking solution 3, it can be dissolved again in a 70 ℃ water bath, shaken well, and then used.

3. All centrifugation steps are performed at room temperature.

4. RNA can inhibit some downstream enzyme reactions, but it will not inhibit PCR. To remove RNA residues, you need to prepare your own RNase A (100 mg/ml) solution.

5. Balance eluent 3 to room temperature.
  

【 Main components 】

Optional reagents:

Lysate 3 (specification 11mL). If the reagent kit is used to extract Omni Swab from Whatman Bioscience, Lysate 3 is not sufficient and needs to be purchased separately.

Customers need to prepare their own items, including: 96~100% ethanol, RNase A (100 mg/ml), PBS, gun head with filter element, metal bath that can be heated to 58 ℃, centrifuge (capable of placing 1.5ml and 2ml centrifuge tubes), vortex oscillator.

Storage conditions and validity period

The reagent kit can be stored for 12 months under dry conditions at room temperature (15-25 ℃). If the room temperature exceeds 25 ℃, the protease and adsorption column 1 can be stored at 2-8 ℃.

Reagent preparation

Before use, add 96-100% ethanol to Lotion 3 and Lotion 4, and add 17ml 96-100% ethanol to Lotion 3 bottles; Add 21ml of 96-100% ethanol to 4 bottles of washing solution. Add 1.5ml of protease solution to the protease bottle for dissolution (store the dissolved protease at 2-8 ℃).

Sample requirements

Oral swab sampling: Firmly adhere the swab to the inner cheek area of the mouth and scrape it 6 times. After collection, air dry the swab for at least 2 hours. Ensure that the person providing the sample has not consumed any food or drink within 30 minutes prior to sample collection.

[Operation Steps]

1. Place the swab into a 2ml centrifuge tube and add 400 μ PBS or 600 μ L (Omni Swab by Whatman Bioscience) PBS and 25 μ L protease, if you want to remove RNA, you also need to add 4 μ L RNase A (100 mg/ml).

2. Add 400 μ L or 600 μ L (Omni Swab from Whatman Bioscience) cracking solution 3 and vortex oscillation mixed for 15 seconds.

Note: Proteinase cannot be directly added to lysate 3.

Take a warm bath at 3.58 ℃ for 10 minutes. Instantaneous centrifugation, plus 400 μ L or 600 μ L (Omni Swab from Whatman Bioscience) 96~100% ethanol, vortex mix well for 15 seconds.

Note: If the room temperature exceeds 25 ° C, cool the ethanol on ice before adding it to the tube.

4. Instantaneous centrifugation, add to the adsorption column, centrifuge at a speed of over 6000g for 1 minute, place the adsorption column in a clean 2 ml collection tube, and discard the collection tube containing the effluent.

Note: 6000g centrifugation will only reduce noise, and centrifugation at a higher speed will not affect DNA extraction. The adsorption column can only add 700 at a time μ If the volume of the solution is relatively large, it can be centrifuged several times on the column.

5. Add 500 μ Centrifuge at a rate of 6000g of washing solution 3 (with ethanol added) for 1 minute, place the adsorption column in a clean 2 ml collection tube, and discard the collection tube containing the effluent.

6. Add 500 μ Centrifuge solution 4 (with ethanol added) at a rate of 20000g for 3 minutes, place the adsorption column in a clean 2 ml collection tube, and discard the collection tube containing the effluent.

7. Suggested steps: Centrifuge at a speed of 20000g for 1 minute.

8. Replace the centrifuge tube and add 100 drops to the middle position of the adsorption membrane μ L-150 μ Elute 3, warm bath at room temperature for 2 minutes, centrifuge at a rate of 6000g for 1 minute.

Note: If the extracted product requires long-term storage, it is recommended to store it at -30 ° C to 15 ° C.