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DNA Purification Kit is suitable for purifying up to 25 μg of high-purity DNA (100 bp-10 kb) from the reaction solution of PCR, enzymatic reaction, and sequencing reaction1. The kit can be stored at room temperature for up to 3 years without showing any reduction in performance and would be stable more than 3 years if stored at 2-8℃.Product Composition
DNA Purification Kit Cat. No. | 5 preps 2101005 | 50 preps 2101050 | 250preps 2101250 |
Spin Columns 2 ml Collection Tubes Buffer P Buffer WB (Concentrate) Buffer TE Instructions | 5 5 3 ml 1.5 ml 0.5 ml 1 copy | 50 50 30 ml 12 ml 5 ml 1 copy | 250 250 75 ml×2 60 ml 25 ml 1 copy |
1. The kit can be stored at room temperature for up to 3 years without showing any reduction in performance and would be stable more than 3 years if stored at 2-8℃.
2. The kit stored at 2-8 °C should be restored to room temperature before use.
The DNA Purification Kit is suitable for purifying up to 25 μg of high-purity DNA (100 bp-10 kb) from the reaction solution of PCR, enzymatic reaction, and sequencing reaction, with a recovery efficiency of 75%~90%, and the purified DNA does not contain primers, enzyme proteins, single nucleotides, fluorescent dyes, or radioisotope-labeled single nucleotides. Suitable for molecular biology experiments with a wide range of requirements.
1. Absolute ethanol
2. 1.5 ml microcentrifuge tube, pipette, and tips
3. Protective equipment such as disposable latex gloves and paper towels
4. Microcentrifuge(s) (with rotor for 1.5 ml and 2 ml microcentrifuge tubes)
6. 3 M sodium acetate (pH 5.0) may be required.
1. If the centrifuge has cooling function, set the temperature to 25 °C.
2. Add absolute ethanol to Buffer WB according to the instructions on the label of the reagent bottle and tick the box on the label to mark "Ethanol added".
1．Add 5 times the volume of Buffer P to 1 volume of the PCR sample or the DNA solution that needs to be cleaned. Do not discard the tip, pipette several times to mix well, transfer the mixture to a spin column (the spin column is placed in a 2 ml collection tube), closed the lid.
* For example, add 500 µl Buffer P for every 100 µl sample.
* If the PCR sample contains mineral oil, it does not need to be removed or included in the sample volume.
* DNA solutions for cleaning include enzymatic reaction solutions (such as enzyme digestion reactions, ligation reactions, etc.), RNase-treated DNA solutions (which can remove degraded RNA) and DNA containing impurities obtained by phenol/chloroform extraction.
* The dye added in Buffer P can indicate the pH. If the mixture turns to violet after adding Buffer P to the sample, add 10 µl of 3M sodium acetate (pH 5.0) and mix, the color of the mixture will turn yellow, otherwise DNA adsorption will be inefficient.
2．Centrifuge at 12000 rpm for 30 s, discard the filtrate and place the spin column back into the collection tube.
3．Add 700 µl Buffer WB to the spin column, closed the lid, centrifuge at 12000 rpm for 30 s. Discard the filtrate and place the spin column back into the collection tube.
* Ensure ethanol has been added into Buffer WB.
4．Centrifuge at 14000 rpm for 1 min.
* If the top speed could not reach 14000 rpm, centrifuge at top speed for 2 minutes.
* Do not pass over this step, or the residual ethanol in the eluted DNA will affects the final applications.
5．Discard the collection tube, place the spin column into a cleaning 1.5 ml centrifuge tube, add 30-50 µl Buffer TE to the center of the column membrane, close the lid, incubate for 1min at room temperature, and centrifuge at 12000 rpm for 30 s.
* The deionized water can also be used to elute DNA, but the pH value should be within 7.0 - 8.5, otherwise it may affect the elution efficiency.
6．Discard the column, the eluted DNA can be used for molecular biology experiment immediately or store the DNA at -20℃ for later use.
