Blood DNA Kit Saliva DNA extraction Kit (column type)

User Manual

Product Name

Common name: Blood DNA Kit Saliva DNA extraction Kit (column type)

Trade name: Blood DNA Kit Saliva DNA extraction Kit (column type)

Model: ZT-004
Intended use

This kit uses a centrifugal adsorption column that can specifically bind nucleic acids and a unique buffer system, which can directly extract DNA (including genomic DNA, viral DNA, Mitochondrial DNA) from saliva samples. The silicon matrix material used in the centrifugal adsorption column can efficiently and specifically adsorb nucleic acids, and combined with a unique buffer system, it can maximally remove impurities such as proteins and other organic compounds in cells. The extracted nucleic acid fragments are complete, highly purified, and of stable and reliable quality. The nucleic acid purified using this kit is suitable for various conventional experiments such as enzyme digestion, PCR, library construction, Southern hybridization, and experiments that require high-quality nucleic acid such as chip hybridization and high-throughput sequencing. This product is expected to be used by professionals, such as technicians and doctors trained in molecular biology technology.

Inspection principle

1. Samples: DNA can be directly extracted from saliva samples (including genomic DNA, viral DNA, Mitochondrial DNA).

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, there is no 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 smaller extracted DNA fragments and a decrease in extraction volume.

If there are precipitates in cracking solution 1 and 3, they can be dissolved again in a 70 ℃ water bath, shaken well, and then used.

3. Balance eluent 1 to room temperature.

4. All centrifugation steps are performed at room temperature.

5. Prepare tissue samples on a cold surface (such as a glass plate, steel plate, or aluminum plate placed above dry ice).

6. If frozen tissue is used, ensure that the frozen sample does not dissolve before adding lysate 1.

7salivaDo not eat, drink, smoke, drink alcohol, or chew gum within 30 minutes before collection. When collecting saliva samples, scrape the upper and lower jaws several times with your tongue, while also scraping the tongue slightly with your teeth.

【 Main components 】

Customers need to prepare their own items, including: 96~100% ethanol, gun head with filter element, metal bath that can be heated to 58 ℃, centrifuge (capable of placing 1.5ml and 2ml centrifuge tubes), and 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 ℃, adsorption column 1 and protease 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 eluent 1 to the protease bottle for dissolution (store the dissolved protease at 2-8 ℃).

Sample Preparation

salivaDo not eat, drink, smoke, drink alcohol, or chew gum within 30 minutes before collection. When collecting saliva samples, scrape the upper and lower jaws several times with your tongue, while also scraping the tongue slightly with your teeth.

[Operation Steps]

1. Take 1.5ml centrifuge tube and add 100 μ Saliva and 100 μ Cracking liquid 1 and 25 μ L protease.

2. Add 200 μ L Cracking liquid 3 is stirred and stirred in a vortex for 15 seconds.

Note: Proteinase cannot be directly added to lysate 3.

Take a warm bath at 3.58 ℃ for 10 minutes.

4. Instantaneous centrifugation, plus 100 μ Mix 96~100% ethanol in a vortex for 15 seconds. Allow to stand at room temperature for 3 minutes.

Attention: If the room temperature exceeds 25 degrees, pre cool the ethanol before use.

5. Instantaneous centrifugation, add 6000g to the adsorption column, centrifuge 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 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.

7. Add 500 μ Centrifuge solution 4 (with ethanol added) at a rate of 6000g for 1 minute, place the adsorption column in a clean 2 ml collection tube, and discard the collection tube containing the effluent.

8. Add 700 μ Centrifuge at a rate of 6000g of 96-100% ethanol for 1 minute, place the adsorption column in a clean 2 ml collection tube, and discard the collection tube containing the effluent.

Centrifuge at a speed of 9.20000 g for 3 minutes.

10. Replace the centrifuge tube with a new one, open the cover and soak at 56 ℃ for 3 minutes until the ethanol evaporates completely.

11. Replace the centrifuge tube and add 20 drops to the middle position of the adsorption membrane μ L-100 μ Eluent 1, warm bath at room temperature for 5 minutes, centrifuge at a rate of 20000g for 1 minute.