Mob/ Whatsapp/Wechat: +86 138 806 76215 | Email: [email protected] / [email protected]
quickly separate and purify genomic DNA from 200-400 µl fresh or frozen anticoagulated whole blood of human or animal within 15-20 minutes. The genomic DNA in the supernatant obtained by centrifugation can be bound to the purification column, and after washing with Buffer WA and Buffer WB to remove the remaining protein and PCR inhibitors on the membrane, the genomic DNA is eluted with Buffer TE, and can be used immediately for various molecular biology experiments including clinical in vitro detection.Product Composition
Blood DNA Mini Kit Cat. No. | 5 preparations 3001005 | 50 preparations 3001050 | 250 preparations 3001250 |
Spin Columns | 5 | 50 | 250 |
2 ml Centrifuge Tubes | 5 | 50 | 250 |
Buffer L1 | 2 ml | 16 ml | 80ml |
Buffer L2 | 2 ml | 16 ml | 80ml |
Buffer WA (concentrate) | 1.9 ml | 12 ml | 60 ml |
Buffer WB (concentrate) | 1.5 ml | 10ml | 50ml |
Buffer TE | 1.2 ml | 12 ml | 60 ml |
Instructions | 1 copy | 1 copy | 1 copy |
If the product is stored at room temperature (0~30°C), the performance of the product will not change significantly within two years; if the product is stored at 2~8°C, the validity period of the product can be extended to more than two years.
This product adopts the national invention patent technology, which can quickly separate and purify genomic DNA from 200-400 µl fresh or frozen anticoagulated whole blood of human or animal within 15-20 minutes. This product does not use proteinase K to digest protein. After the whole blood is dissolved in Buffer L1, the hemoglobin is removed by precipitation in Buffer L2. The genomic DNA in the supernatant obtained by centrifugation can be bound to the purification column, and after washing with Buffer WA and Buffer WB to remove the remaining protein and PCR inhibitors on the membrane, the genomic DNA is eluted with Buffer TE, and can be used immediately for various molecular biology experiments including clinical in vitro detection.
Equipment and Reagents to Be Supplied by User
1. Absolute ethanol
2. 1.5 ml centrifuge tubes
3. Pipettes and tips (in order to avoid cross-contamination between samples, it is recommended to use pipette tips with filter elements)
4. Protective equipment such as latex gloves, disposable masks, and paper towels
5. Microcentrifuge(s) (with rotor for 1.5 ml and 2 ml microcentrifuge tubes)
6. Vortexer, Water bath
7. normal saline maybe needed
1. If the centrifuge has refrigeration function, please set the temperature to 25°C.
2. Incubate Buffer TE at 56°C (can be omitted).
3. Add absolute ethanol to Buffer WA and Buffer WB according to the instructions on the label of the reagent bottle, and tick the box on the label to mark "Alcohol added".
The following protocol is designed for 400 µl blood sample, reduce the amount of Buffer L1 and Buffer L2 in proportion if the volume of blood is between 200 - 400 µl, adjust the volume to 200 µl with normal saline if the volume of blood is less than 200 µl (Buffer L1: blood : Buffer L2 = 3 : 4 : 3 must be strictly followed, otherwise the subsequent steps will not be performed), and the amount of other reagents remains unchanged.
1. Add 300 µl Buffer L1 in a 1.5 ml microcentrifuge tube (not provided).
2. Add 400 μl anticoagulated blood. Close the lid, vigorously shake microcentrifuge tube 3 - 5 times, and then mix by vortex for 30 seconds.
3. Add 300 μl Buffer L2. Close the lid, vigorously shake microcentrifuge tube 3 - 5 times, and then mix by vortex for 30 seconds.
* A large amount of hemoglobin precipitation will appear in this step.
4. Centrifuge at 13000 rpm for 2 minutes.
5. Placed a spin column in a 2 ml collection tube, pour the supernatant from step 4 into the spin column, close the lid, and centrifuge at 12000 rpm for 30 seconds.
6. Discard the filtrate. Place the spin column back into the 2 ml collection tube. Add 500 μl Buffer WA containing ethanol, close the lid and centrifuge at 12000 rpm for 30 seconds.
* Ensure ethanol has been added into Buffer WA.
7. Discard the filtrate. Place the spin column back into the 2 ml collection tube. Add 600 μl Buffer WB containing ethanol, close the lid and centrifuge at 12000 rpm for 30 seconds.
* Ensure ethanol has been added into Buffer WB.
8. Discard the filtrate. Place the spin column back into the 2 ml collection tube. Centrifuge at 14000 rpm for 1 minute.
* If the full speed could not reach 14,000 rpm, centrifuge at full speed for 2 minutes.
* Do not omit this step, otherwise, it may cause problems in downstream applications due to the residual ethanol in the eluate.
9. Discard the 2 ml collection tube. Place the spin column into a clean 1.5 ml microcentrifuge tube (not provided). Add 100 - 200 μl pre-warmed Buffer TE to the center of the membrane. Close the lid, incubate for 1 minute at room temperature, and then centrifuge at 12000 rpm for 30 seconds.
* Recovery may be increased by 10 - 20 % if incubate longer (5 - 15 minutes) after Buffer TE addition.
10. Discard the spin column. Eluted DNA can be used in downstream applications immediately, or store at -20℃ for later use.
