Products

Blood gDNA Extraction Mini Kit(SpinColumn)

Packing Specification：

Product Introduction:
The unique Binding Buffer/Proteinase K rapidly lyses cells and inactivates intracellular nucleases. Genomic DNA then selectively binds to the silica matrix membrane in the spin column under high chaotropic salt conditions. Through a series of rapid washing-centrifugation steps, impurities such as cellular metabolites and proteins are removed by the Inhibitor Removal Buffer and Wash Buffer. Finally, pure genomic DNA is eluted from the silica matrix membrane using a low-salt elution buffer.

Product Features:
1.Eliminates the need for toxic reagents such as phenol and ethanol precipitation steps.
2.Rapid and simple: Processing of a single sample can generally be completed within 20 minutes.
3.Multiple column washing steps ensure high purity. Typically, 3-12 µg of genomic DNA can be extracted from 200 µL of whole blood, with a typical OD260/OD280 ratio of 1.7～1.9 and a length of 30 kb -50 kb. It can be directly used for PCR, Southern blot, and various restriction digestion reactions.
4.Optimized red blood cell lysis buffer formula (available for separate purchase) enables rapid and complete lysis.

Application Scope:
Suitable for the rapid extraction of genomic DNA from whole blood.

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.Take 200 µL of fresh, frozen, or anticoagulated blood and place it into a 1.5 mL centrifuge tube.
If the initial volume of whole blood is less than 200 µL, make up to 200 µL with Buffer BB.If the initial volume is 200 µL-300 µL, increase reagent dosages proportionally in subsequent steps.If the initial volume is 300 µL-1 mL, perform red blood cell lysis first (see Appendix at the end of this manual).
For anticoagulated blood from avians, amphibians, or lower organisms (with nucleated red blood cells), use 5-20 µL of blood and make up to 200 µL with Buffer BB before proceeding.

2.Add 20 µL of Proteinase K (20 mg/mL) solution, mix thoroughly. Add 200 µL of Binding Buffer CB, vortex immediately to mix well, and incubate at 70℃ for 10 minutes. The solution should become clear (but dark in color).
Optional Step (generally not required): To remove residual RNA, add 20 µL of RNase A (25 mg/mL) solution before adding 200 µL of Binding Buffer CB, vortex to mix, and incubate at room temperature for 5-10 minutes.

Pretreat the adsorption column with Equilibration Buffer (mandatory step; refer to "About the Use of Equilibration Buffer" above).
3.After cooling, add 100 µL of isopropanol, vortex immediately to mix well (flocculent precipitation may form).
Thorough mixing at this step is critical; insufficient mixing will severely reduce yield. Vortex for 15 seconds if the sample is viscous and difficult to mix.

4.Load the mixture (including any precipitation) into a Spin AC Adsorption Column (placed in a collection tube). Centrifuge at 13,000 rpm for 30-60 seconds and discard the waste liquid in the collection tube.

5.Add 500 µL of Inhibitor Removal Buffer IR, centrifuge at 12,000 rpm for 30 seconds, and discard the waste liquid.

6.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.

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

8.Place the Spin AC Adsorption Column back into the empty collection tube. Centrifuge at 13,000 rpm for 2 minutes to completely remove residual Wash Buffer (residual ethanol may inhibit downstream reactions).

9.Transfer the Spin AC Adsorption Column to a clean centrifuge tube. Add 100 µL of Elution Buffer EB (preheating in a 65-70℃ water bath can improve yield) to the center of the adsorption membrane. Incubate at room temperature for 3-5 minutes, then centrifuge at 12,000 rpm for 1 minute. Reapply the eluate to the adsorption column, incubate at room temperature for 2 minutes, and centrifuge at 12,000 rpm 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 50 µL (excessively small volume reduces DNA elution efficiency and yield).

10.Store DNA at 2-8℃ for short-term use or at -20℃ for long-term storage.