StarPure Gel Recovery Kit (Membrane Column Method)

• High recycling efficiency
• The recovered DNA is of high purity

• PCR product recovery
• Enzyme digestion product recovery
• Other applications of agarose gel electrophoresis for DNA separation

Since DNA may be lost during electrophoresis, in order to determine the actual recovery efficiency of kits from different manufacturers, we cut 100 mg of clean blank gel and placed it in a 1.5 mL EP tube.
At the same time, 1000 ng of 450 bp DNA fragments were added, and the gel dissolution and subsequent DNA recovery processes were performed using kits from different manufacturers (according to the respective manufacturers' instructions).
The DNA recovery rate of the Qihengxing membrane column gel extraction kit was higher than that of supplier A and supplier B (Fig. 1).


Fig.1 DNA recovery rate of gel recovery kits from different manufacturers

Using a DNA marker (B500351) from Bioengineering as a sample, 20 μL of sample was loaded and separated by electrophoresis in a 1% gel. The fragments were manually cut out from the agarose gel.
DNA fragments of 5000 bp, 1000 bp, and 500 bp were recovered using the StarPure gel recovery kit (column method) and eluted with 30 μL; the elution product of each fragment
The samples (5 μL) and marker (3 μL) were re-run on agarose gel electrophoresis and the results were compared.


Fig.2 Electrophoresis of recovered DNA fragments

Notes:

1. Materials to prepare: centrifuge, water bath (or metal bath), isopropanol, 1.5 mL centrifuge tubes, and 80% ethanol.
2. StarPure Gel Buffer A may produce precipitation at low temperatures. Please check before use. If precipitation is present, dissolve it at 37°C and cool to room temperature before use.

3. Before first use, add isopropyl alcohol to the StarPure Gel Washing Buffer C bottle (according to the volume indicated on the bottle label) and mix thoroughly.

4. Please adjust the water bath (or metal bath) to 56°C in advance.

Before use, add isopropyl alcohol (according to the label on the bottle) to StarPure Gel Washing Buffer C.

1. Separate the target DNA fragment from other fragments by agarose gel electrophoresis. Use a clean scalpel blade to cut the agarose gel containing the target DNA, place it in a 1.5 mL centrifuge tube, and weigh it (usually the gel block cut from each band should be within 100 mg. Please cut away as much blank gel block as possible).
2. According to the weight of the gel block, add 250 μL of StarPure Gel Buffer A for every 100 mg of agarose gel block (if the gel block is less than 100 mg, it will still be calculated as 100 mg);
3. Place the centrifuge tube containing the gel block in a 56°C water bath for 5 minutes until the gel block is completely dissolved;
4. Remove the centrifuge tube, equilibrate to room temperature, add 150 μL of isopropanol, and shake to mix;
5. Add the solution obtained in step 4 to the DNA adsorption column (the adsorption column is placed in the collection tube) and let it stand at room temperature for 5-10 minutes;
6. Centrifuge at 12000 rpm for 2 min and discard the waste liquid in the collection tube.
7. Add 600 μL StarPure Gel Washing Buffer C (make sure to add an equal volume of isopropanol before use), shake to mix, centrifuge at 12000 rpm for 2 min, and discard the waste liquid in the collection tube.
8. Add 600 μL of 80% ethanol, centrifuge at 12,000 rpm for 2 min, and discard the waste liquid in the collection tube.
9. Centrifuge at 12000 rpm for 2 min to remove residual ethanol.
10. Place the adsorption column in a new collection tube, add 50-100 μL of elution buffer (TE or deionized water) to the middle part of the adsorption membrane, leave it at room temperature for 2-5 minutes, centrifuge at 12,000 rpm for 2 minutes, collect the plasmid solution into a centrifuge tube, and store the DNA at -20℃ or use it immediately.