01/Product introduction

This kit is based on the principle of silicon-based magnetic beads to specifically adsorb DNA, enzymatically releases blood genome, binds DNA molecules in a specific high-efficiency lysis buffer system, changes the environmental ionic strength, and realizes the separation and purification of genomic DNA. It is suitable for efficiently extracting up to 50kb genomic DNA from saliva or buccal swabs. An excellent buffer system can remove impurities such as glycosaminoglycans and viscous proteins in saliva to the greatest extent. The obtained genomic DNA is suitable for PCR reactions, enzyme digestion reactions, Library construction and Southern hybridization and other downstream experiments. It can be operated manually by the operator, and it is also suitable for high-throughput automatic operation.

02/Product advantage

1. The operation process is safe and non-toxic

2. Compatible with genome extraction of saliva and nasal swab samples at the same time

3. Easy and quick operation, good quality

4. Suitable for hand-held and high-throughput

03/Product content

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 a corresponding volume of absolute ethanol to BufferSW1 and BufferSW2.

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

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

07/How to use

1. Take 500ul saliva in a 1.5ml clean centrifuge tube.

2. Add 300ul Buffer KB and 20ul Proteinase K to it, vortex and mix for 30 seconds.

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

4. Add 200ul of isopropanol and vortex for 20s (note: there may be agglomeration at this time, must be fully dispersed), add 20ul of magnetic bead suspension (note: vortex and mix the magnetic bead suspension before adding).

5. Vortex and mix for 30 seconds, and let stand for 1 minute.

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 SW1, vortex and mix for 2 minutes, perform magnetic separation, and remove the liquid other than the magnetic beads (Caution: Do not suck off the magnetic beads).

9. Repeat step 8 once.

10. Remove the centrifuge tube from the magnetic stand, add 600ul Buffer SW2, vortex and mix for 2 minutes, perform magnetic separation, and remove the liquid other than the magnetic beads (Caution: Do not suck off 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 the genome quality).

13. Remove the centrifuge tube, add 50-200ul Buffer STE, pipette and mix 20 times, incubate at 65°C for 8-10 minutes, and mix twice during the period.

14. The transfer centrifuge tube is placed on a magnetic stand for magnetic separation, and the liquid except the magnetic beads is transferred to a new centrifuge tube and stored 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 Buffer SW1 and Buffer SW2 by 100ul.

2. Use fresh saliva 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. Before eluting with Buffer STE, it was not fully dried, 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 Buffer STE 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 magnetic beads.

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

5. If the drying time exceeds 10 minutes, the magnetic beads are too dry and the elution efficiency is reduced.

6. In the process of elution with Buffer STE, the magnetic bead clumps were not sufficiently dispersed, resulting in insufficient genome release.