The free DNA (cfDNA) kit mainly uses the magnetic bead method. The detection principle is that the magnetic beads (mainly ferrite) can be quickly captured by the magnetic beads to form a magnetic bead complex through copolymerization and surface modification of free DNA, and settle down from the liquid phase under the action of a magnetic field. After simple washing, separation, changing washing and cleaning conditions, etc. Free DNA bound to the surface of the magnetic beads is eluted.

The kit provides a simple, fast and efficient CFDNA (Free circulating/Cell-free DNA) extraction method, which is suitable for extracting CFDNA from serum, plasma, lymph, urine and other cell-free body fluids. The purified CFDNA is of stable and reliable quality and can be used for downstream routine experiments. The kit can be used in conjunction with the nucleic acid extractor of the transfer liquid method to perform large-scale extraction simply and quickly, which greatly reduces the workload of the experimenter and the human error in the experiment.The free DNA (cfDNA) kit mainly uses the magnetic bead method. The detection principle is that the magnetic beads (mainly ferrite) can be quickly captured by the magnetic beads to form a magnetic bead complex through copolymerization and surface modification of free DNA, and settle down from the liquid phase under the action of a magnetic field. After simple washing, separation, changing washing and cleaning conditions, etc. Free DNA bound to the surface of the magnetic beads is eluted.

The kit provides a simple, fast and efficient CFDNA (Free circulating/Cell-free DNA) extraction method, which is suitable for extracting CFDNA from serum, plasma, lymph, urine and other cell-free body fluids. The purified CFDNA is of stable and reliable quality and can be used for downstream routine experiments. The kit can be used in conjunction with the nucleic acid extractor of the transfer liquid method to perform large-scale extraction simply and quickly, which greatly reduces the workload of the experimenter and the human error in the experiment.

•Using the patented technology of magnetic bead-based classification and screening to ensure the effective extraction of cfDNA, the fragment length is mainly 170bp

•Fast and direct experiment process, the whole process only takes 2 hours. At the same time, there is no need to run gel, column or centrifugation operation

•The product has high purity, effectively avoiding protein, salt, nuclease, PCR inhibitor and other impurities

• Sensitive and efficient DNA capture technology to ensure a minimum of 80% DNA extraction efficiency

• Flexible experimental design requirements, suitable for high-throughput research01/Product introduction

Free DNA, also known as free circulating nucleic acid, refers to DNA that is free from cells in the circulating blood.

Often no more than 800Bp, mainly in serum and plasma. Free DNA fragments are used in the early diagnosis, prognosis, and monitoring of diseases.

It has important potential value in aspects such as measurement. This kit is specially designed for the extraction of free DNA, based on the specific absorption of silicon-based magnetic beads.

Attached to the principle of DNA, it can achieve impurity removal and enrichment in various extraction components, and obtain pure and high-quality free DNA molecules, suitable for

It is used for downstream molecular operations such as PCR reaction, qPCR detection, and next-generation sequencing (NGS).

02/Product advantage

1. Strong universality, suitable for the extraction of fresh or frozen anticoagulated serum, plasma and cell-free body fluid samples

2. Stable performance, non-toxic extraction process

3. High quality and good purity

04/Storage conditions

It is valid for 1 year when stored at room temperature.

05/Bring your own reagents

Isopropanol, absolute ethanol

06/Notes

1. Before using for the first time, add the corresponding volume of absolute ethanol to BufferCW1 and BufferCW2.

2. Shake the solution in each bottle before each use to make the system uniform and ensure the extraction effect.

3. The sample should avoid repeated freezing and thawing (generally no more than 3 times), otherwise the extraction quality will be reduced.

4. Samples can be stored at 4°C for a week for short-term storage, and for long-term storage at -80°C.

5. If precipitation occurs after the solution is left for a period of time, it can be dissolved in a 37°C water bath without affecting the effect.

6. Use sterile centrifuge tubes and pipette tips to avoid contamination by exogenous nucleases.

07/How to use

1. Take 600ul sample in a 1.5ml clean centrifuge tube.

2. Add 400ul Buffer KD and 20ul Proteinase K to it, vortex and mix for 30 seconds.

3. Incubate at 68°C (both in water bath and metal bath) for 10 minutes, vortex and mix for 5 seconds every 2-3 minutes.

4. Add 200ul isopropanol and vortex for 20s, add 20ul magnetic bead suspension (note: vortex and mix magnetic bead suspension before adding).

5. Vortex and mix for 30smin, and let stand for 1min.

6. Repeat step 5 twice.

7. Place the transfer centrifuge tube on the magnetic stand. After magnetic separation, carefully suck off the liquid (Caution: Do not suck off the magnetic beads).

8. Remove the centrifuge tube from the magnetic stand, add 800ul Buffer CW1, vortex and mix for 2 minutes, perform magnetic separation, and remove by suction.

Liquid other than magnetic beads (Caution: Do not suck away the magnetic beads).

9. Repeat step 8 once.

10. Remove the centrifuge tube from the magnetic stand, add 600ul Buffer CW2, vortex and mix for 2 minutes, perform magnetic separation, and remove by suction.

Liquid other than magnetic beads (Caution: Do not suck away the magnetic beads).

11. Repeat step 10 once.

12. Transfer the centrifuge tube to a magnetic rack and dry it at room temperature for 8-10 minutes (Note: the drying time should not exceed 10 minutes, otherwise it will

Affect genome quality).

13. Take off the centrifuge tube, add 80-150ul BufferCTE, pipette and mix 20 times, incubate at 65°C for 8 min.

Mix by blowing and sucking twice.

14. The transfer centrifuge tube is placed on the magnetic stand for magnetic separation, and the liquid except the magnetic beads is transferred to a new centrifuge tube.

Store at -20°C.

08/Common problems and coping strategies

◆The purity of genomic DNA obtained is low

1. It is recommended to increase the elution volume of BufferCW1 and BufferCW2 by 100ul.

2. Use fresh samples for extraction.

3. During the washing process, the magnetic beads were not sufficiently dispersed, and there was protein aggregation. It is recommended to increase the strength and time of blowing or vortex mixing.

4. It was not fully dried before elution with BufferCTE, and a small amount of ethanol remained.

◆Low concentration of genomic DNA obtained

1. It is recommended to replace with fresh samples for extraction.

2. Reduce the amount of BufferCTE, or extend the incubation time at 65°C to 15min.

3. During the elution process, if the magnetic beads are not sufficiently magnetically separated, the liquid will be transferred, resulting in the loss of the magnetic beads.

4. Increase the magnetic beads by 5-10ul to increase the adsorption strength, thereby increasing the yield.

5. The drying time exceeds 10 minutes, the magnetic beads are too dry, and the elution efficiency decreases.

6. In the process of elution with BufferTE, the magnetic bead clumps were not sufficiently broken up, resulting in insufficient genome release.

◆Agglomeration of magnetic beads occurred during the extraction process

In the magnetic bead extraction process, because a large amount of chromatin nucleic acid is adsorbed on the surface of the magnetic beads, magnetic bead knots will appear in most cases
 
Group phenomenon. This phenomenon can be used as an indicator of nucleic acid content. Generally speaking, clumping indicates that the sample is good and the adsorption process

Smooth and efficient, as long as the operator fully breaks up the clumps during the washing and elution process, an ideal genomic DNA fraction can be obtained

son.