Products

PCR Product DNA Purification Kit(SpinColumn)

Packing Specification：

Product Introduction:
In the presence of high chaotropic salts, DNA fragments selectively bind to the silica matrix membrane in the spin column. Through a series of rapid washing-centrifugation steps, impurities such as primers, nucleotides, proteins, and enzymes are removed by deproteinization solution and wash buffer. Finally, pure DNA is eluted from the silica matrix membrane using a low-salt and high-pH elution buffer.

Product Features:
1.The silica matrix membranes in the spin columns are all high-quality specially manufactured adsorption membranes, ensuring minimal variation in adsorption capacity between columns.
2.Uses high-quality binding buffer without sodium iodide or perchlorate found in traditional binding buffers, which do not inhibit downstream reactions such as restriction digestion, ligation, and cloning after recovery.
3.Binding Buffer BB is formulated with phenol red to appear yellow, facilitating pH monitoring for optimal binding efficiency, greatly improving recovery rate.
4.Rapid and convenient: Eliminates the need for toxic reagents such as phenol and chloroform, as well as ethanol precipitation.

Application Scope:
Suitable for purification and recovery of PCR reaction products, digested DNA fragments, and probe labeling, as well as concentration of DNA samples.

Operation Steps:
Tip: Before first use, add the specified volume of absolute ethanol to the Wash Buffer WB bottle, mix thoroughly. Mark the corresponding box immediately after adding ethanol to avoid repeated additions!

1.Add 500 μL of Binding Buffer BB to every 100 μL of PCR amplification product or digested sample, and mix thoroughly. (If the initial volume is less than 100 μL, adjust to 100 μL with double-distilled water in advance.)
Pretreat the adsorption column with Equilibration Buffer (mandatory step; refer to "About the Use of Equilibration Buffer" above for specific methods.)

2.Load the solution obtained in the previous step into the Spin EC Adsorption Column (placed in a collection tube), let stand at room temperature for 1 minute, centrifuge at 12,000 rpm for 30-60 seconds, and discard the waste liquid in the collection tube.
After mixing the filtered binding buffer with the residual strongly alkaline Equilibration Buffer in the collection tube, the solution may change color from yellow to orange-red or even purple, which is a normal color change of the phenol red pH indicator under alkaline conditions.

3.Add 600 μL of Wash Buffer WB (check if absolute ethanol has been added!). Centrifuge at 12,000 rpm for 30 seconds and discard the waste liquid.

4.Add another 600 μL of Wash Buffer WB, centrifuge at 12,000 rpm for 30 seconds, and discard the waste liquid.

5.Place the Spin EC Adsorption Column back into the empty collection tube. Centrifuge at 12,000 rpm for 2 minutes to completely remove residual wash buffer (residual ethanol may inhibit downstream reactions).

6.Transfer the Spin EC Adsorption Column to a clean centrifuge tube. Add 50 μL of Elution Buffer EB (preheating the elution buffer in a 65-70℃ water bath can improve yield) to the center of the adsorption membrane. Incubate at room temperature for 2 minutes, then centrifuge at 12,000 rpm for 1 minute. For larger DNA yield, reapply the eluate to the adsorption column and centrifuge for 1 minute.
Higher elution volume leads to higher elution efficiency. To obtain higher DNA concentration, the elution volume can be appropriately reduced, but the minimum volume should not be less than 25 μL (excessively small volume reduces DNA elution efficiency and yield).