Mag TR DNA Kit

TR DNA Kit is based on self-developed high-efficiency separation magnetic beads. With the cooperation of a unique buffer system, it can extract trace DNA fragments of host cells (CHO, Vero, NS0, MDCK, E.Coli, etc.) from biological product samples less than 200 µl , the purified nucleic acid can be applied to various routine operations, including PCR, RT-PCR, fluorescent quantitative PCR and other downstream experiments.

Components

Storage and stability

The kit can be stored at room temperature for 12 months.

Required instruments and consumables:

1. Nuclease-free tips and 1.5 ml , Nuclease-free centrifuge tubes

2. High-speed centrifuge

3. Constant temperature metal bath

4. Magnetic stand

Sample Preprocessing:

1. Take a sterile 1.5 ml centrifuge tube, add 20 µl Proteinase K, 125 µl Buffer MDA, 275 µl

Buffer GAL, and then add 200 µl of the sample to be tested (real sample/NEC/ERC).

2. Cap tubes tightly and vortex briefly, then centrifuge briefly.

3. Incubate the centrifuge tube at 56°C for 30 min.

DNA Extraction:

1. After the pretreatment is completed, take out the centrifuge tube and let it stand at room temperature, then centrifuge briefly to collect the liquid to the bottom of the tube.

2. Add 400 μl Buffer IP and 20μl Magnetic beads C to the centrifuge tube, close the cap tightly, and vortex at room temperature to mix 5 min. Place the centrifuge tube on the magnetic stand and let it stand for 30 sec. After the magnetic beads are completely absorbed, carefully suck off the liquid.

3. Remove the centrifuge tube from the magnetic stand, add 900μl Buffer MW, shake and mix for 30 sec. Place the centrifuge tube on the magnetic Let it stand on the rack for 30 sec. After the magnetic beads are completely absorbed, carefully suck off the liquid.

4. Remove the centrifuge tube from the magnetic stand, add 900μl Buffer MWP, shake and mix for 30 sec. Place the centrifuge tube in the magnetic Let stand on the force rack for 30 sec. After the magnetic beads are completely absorbed, carefully suck off the liquid.

5. Keep the centrifuge tube on the magnetic stand, open the tube cap, and dry at room temperature for 3 minutes.

6. Remove the centrifuge tube from the magnetic stand, add 50μl Nuclease-free H2O, and incubate at 70°C for 5 minutes.

7. Place the centrifuge tube on the magnetic stand and let it stand for 1 min. After the magnetic beads are completely absorbed, carefully transfer the nucleic acid solution to a Store in a new centrifuge tube (self-prepared) and in appropriate conditions

Matters Need Attention

1. Please read the instructions for use carefully before use, and operate in strict accordance with the instructions for use.

2. Samples should avoid repeated freezing and thawing, otherwise the amount of nucleic acid extraction will be reduced.

3. When using this kit to extract DNA,remember to wear disposable latex gloves and masks, and pay special attention to prevent nucleic acid degradation during the operation.

4. To ensure that the experimental results are credible:It is recommended to add extraction  recovery rate quality control (ERC) samples, and perform nucleic acid extraction and     quantitative detection steps simultaneously to determine the efficiency of nucleic acid    extraction and recovery.

It is recommended to add negative quality control (NCS) samples, and perform nucleic acid extraction and quantitative detection steps simultaneously to determine whether there

is contamination in the nucleic acid extraction process.

The recommended sample preparation method is as follows:

(a) Add sample and recover quality control ERC sample: add 20 µl host DNA standard to 180 µl sample to be tested and mix well as ERC;

(b) Negative quality control NCS sample: 200 µl 1 × TE buffer (DNA diluent or basic solvent for biological products), as NCS.