StarPure Ultra-Fast DNA Methylation Conversion Kit (Column-Based)

The StarPure Ultra-Fast DNA Bisulfite Conversion Kit (column-based) can rapidly convert unmethylated cytosine in DNA samples to uracil, while maintaining methylated cytosine. After high-temperature bisulfite treatment, double-stranded DNA melts into single strands. Under the action of HSO₃⁻, cytosine (C) residues undergo deamination and are converted to uracil (U), while methylated residues remain unchanged. Uracil (U) is then replaced by thymine (T) during the subsequent PCR amplification process. The conversion reaction takes only 10 minutes, and the use of in-column desulfurization technology simplifies the operation process. The DNA input range is 2 ng to 2 μg, and the conversion efficiency of unmethylated cytosine is ≥99%. The converted products are suitable for downstream applications such as PCR amplification and NGS sequencing.

1. You need to prepare anhydrous ethanol before use.
2. This kit can process approximately 2 to 2000 ng of DNA sample, and the recommended input amount of gDNA is greater than 200 ng.
3. To avoid reagent crystallization affecting the experiment, please store the kit at room temperature.
4. It is recommended that all reagents be incubated in a 37°C oven/water bath for 30 min before the experiment to ensure that the solution is free of crystals.
5. DNA treated with this kit is not suitable for quantification by OD260 or determination of sample quality by OD260/OD280.
6. Ensure that the reagents in the kit are fully mixed during the experiment and close the lid immediately after use to prevent the reagents from being exposed to air for a long time, which may cause volatilization, oxidation, and pH changes.
7. The shelf life of DB solution containing anhydrous ethanol is 14 days. The formation of a white precipitate is normal due to the nature of the reagent. Be careful not to absorb the precipitate during use. Prepare the solution immediately before use, adding 4 times the volume of anhydrous ethanol each time.

Product components


Storage and transportation conditions

This product should be stored and transported at room temperature (15~25℃)

1. Methylation conversion

1.1. Place 5 μL of the DNA to be transformed into a 200 μL PCR tube (if the DNA volume is less than 5 μL, it is recommended to use RNase-free H2O to make up to 5 μL). Add 9 volumes of Transformation Buffer BS and vortex to mix thoroughly (the total volume is recommended not to exceed 100 μL).

Place the PCR tubes from step 1.1 in a PCR instrument and run the following program:

2. Product purification

2.1. Transfer the transformed product to a clean 2 mL centrifuge tube. Add 5 times the total volume of the liquid in the centrifuge tube (250 μL) of Binding Buffer BB and vortex to mix.

2.2. Place the purification column in the collection tube and transfer the solution obtained in step 1 completely to the purification column. Let it stand at room temperature for 2 minutes. Centrifuge at 25°C and 12,000 rpm for 1 minute. Discard the filtrate and return the purification column to the collection tube.

2.3. Add 600 μL of washing buffer (WB) to the purification column and centrifuge at 12,000 rpm at 25°C for 1 min. Discard the filtrate and return the purification column to the collection tube.

2.4. Add 600 μL of desulfurization buffer DB to the purification column, let it stand at room temperature for 15 minutes, centrifuge at 25°C and 12,000 rpm for 1 minute, discard the filtrate, and return the purification column to the collection tube;

2.5. Add 600 μL of washing buffer (WB) to the purification column. Centrifuge at 12,000 rpm at 25°C for 1 min. Discard the filtrate and return the purification column to the collection tube.

2.6. Repeat step 2.5 twice;

2.7. Centrifuge the empty purification column at 12,000 rpm at 25°C for 3 min. Transfer the column to a clean 1.5 mL centrifuge tube and air-dry at room temperature with the lid open for 3 min.

2.8. Add 50 μL of eluent (TE or deionized water) to the center of the purification column membrane. Let stand at room temperature for 2 minutes. Centrifuge at 12,000 rpm at 25°C for 2 minutes. Collect the product solution, which is the purified transformed DNA product.

2.9. The collected DNA products can be stored at 4°C for short-term storage and at -20°C for long-term storage.