Blood DNA Kit Genome DNA extraction kit for dry blood spot (column type)

User Manual

Product Name

Common name: Blood DNA Kit Genome DNA extraction kit for dry blood spot (column extraction type)

Trade name: Blood DNA Kit Genome DNA extraction kit for dried blood spot (column extraction type)
Model: ZT-008

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 dry blood spots (anticoagulant or non anticoagulant) on filter paper. 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. Sample: DNA (including genomic DNA, viral DNA, Mitochondrial DNA) can be directly extracted from the dry blood spot (anticoagulant or non anticoagulant) on the filter paper.

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 3 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.

【 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 ° C/85 ° C/70 ℃ and can be mixed evenly, centrifuge (which can fit 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 protease solution to the protease bottle for dissolution (store the dissolved protease at 2-8 ℃).
  

[Operation Steps]

1. Use a Hole punch to cut three discs with a diameter of 3mm from the dry blood spot filter paper, put them into a 1.5ml centrifuge tube, and add 180 μ Cracking liquid 1.

Warm bath at 2.85 ° C for 10 minutes.

3. Instantaneous centrifugation, plus 20 μ L protease, thoroughly mix well, mix well at 58 ℃ and take a warm bath for 1 hour.

4. Instantaneous centrifugation, adding 200 μ L cracking solution 3, thoroughly mix well, mix well at 70 ℃ and take a warm bath for 10 minutes.

5. Instantaneous centrifugation, adding 200 μ Ethanol (96-100%), cover and mix thoroughly.

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

6. 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.

7. 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.

8. 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.

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

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

Note: Three 3mm diameter discs can extract approximately 150ng of DNA if anticoagulant, and approximately 75ng of DNA if non anticoagulant. If the yield of non anticoagulant extraction is insufficient, the number of discs can be increased to 6. When extracting products for PCR amplification, the amount of template used should not exceed 10% of the total volume of the reaction system.