4. Tissue/Cell/Blood Genomic DNA Extraction Kit, 50T $149

Product Information

Product Description/Introduction

This kit releases genomic DNA from tissue cells through a specially optimized lysis buffer, and then purifies genomic DNA through a specific DNA-binding membrane. It is suitable for isolating genomic DNA from animal tissues, cultured cells and whole blood. 10-30 μg high purity and complete genomic DNA can be rapidly extracted and purified from 2-30 mg animal tissues, 106-107 freshly cultured cell suspensions and 5-100 μL whole blood of non-nucleated or nucleated red cells (including anticoagulant). Genomic DNA obtained is ready for a variety of applications, such as PCR, Southern blotting, RAPD, AFLP, RFLP and other molecular experiments.

Storage and Shipping Conditions

Proteinase K and RNase A should be shipped with wet ice, and stored at -20℃; Other reagents are shipped and stored at room temperature for up to 1 year.

Product Contents

Before starting (please read carefully)

1. If a precipitate has formed in Buffer GA, heat it at 65°C until the precipitate has fully dissolved.

2. Before first use, add 18 mL of absolute ethanol to Buffer PD, add 56 mL of absolute ethanol to Buffer PW and mix well separately.

Assay Protocol / Procedures

1. Samples Preparation

a. Take out 2-30 mg of fresh or frozen tissue at -80°C, place them in a 1.5 mL centrifuge tube or special grinding tube containing 2-3 grinding beads with a diameter of 3 mm (G0203-150G recommended), add 200 μL of Buffer GA, and use a Tissue Homogenizer (KZ-5F-3D recommended) to thoroughly grind the tissue to homogenization at room temperature (if the tissue is not thoroughly homogenized, it will affect the yield and quality of DNA).

b. Take 106-107 of cell suspension into a 1.5 mL centrifuge tube, centrifuge at 1,000 rpm for 5 minutes, and discard supernatant. Add 200 μL Buffer GA, mix well by votex.

c. Take 50-100 μL whole blood of non-nucleated erythrocytes (including anticoagulant) into a 1.5 mL centrifuge tube, add Buffer GA solution to 200 μL, mix well. with votex.

Note: For processing a larger volume of blood, add 3 times volume of erythrocyte lysate to the sample and mix it upside down, let it stand at room temperature for 5 min, mix it upside down several times during incubation, Centrifuge it at 3,000 rpm for 10 min, gently remove the supernatant with pipette to leave the white blood cell precipitate, add 200 μL Buffer GA, mix well with votex.

d. Take 5-20 μL whole blood of nucleated erythrocytes (including anticoagulant) into a 1.5 mL centrifuge tube, add Buffer GA solution to 200 μL, mix well with votex.

2. Add 20 μL Proteinase K and 4 μL of RNase A to the sample from step 1, incubate at 56℃ for 30 minutes (Mix every 10 minutes to accelerates tissue lysis).

Note: Please extend time for materials that are difficult to lysis.

3. Centrifuge at 12,000 rpm for 2 min, transfer the tissue supernatant to a Nuclease-free centrifuge tube, avoiding aspirating the pellet. (Please skip this step if the sample is cell or blood).

4. Add 200 μL of Buffer GB to the centrifuge tube, mix upside and down, and incubate at 70 °C for 10 min.

5. Add 200 μL of absolute ethanol to the centrifuge tube, mix upside and down (any visible precipitate that may form after adding isopropanol), and then centrifuge at 12,000 rpm for 2 min

6. Transfer the supernatant to a Nuclease-free centrifuge tube, avoiding aspirating the pellet.

7. Place the DNA Spin Columns into the Collection Tubes, transfer all the mixture to the DNA Spin Columns.

8. Centrifuge at 12,000 rpm for 30 seconds, discard the flow-through and put the DNA Spin Columns back into the collection tube.

9. Add 500 μL Buffer PD to the DNA Spin Columns, centrifuge at 12,000 rpm for 30 seconds, discard the flow-through and put the DNA Spin Columns back into the collection tube.

10. Add 600 μL Buffer PW to the DNA Spin Columns, centrifuge at 12,000 rpm for 30 seconds, discard the flow-through and put the DNA Spin Columns back into the collection tube.

11. Repeat Step 10.

12. Centrifuge at 12,000 rpm for 2 minutes to remove any residue.

13. Put the DNA Spin Columns into a new centrifuge tube, stand at room temperature for 5-10 minites.

14. Add 50-100 μL Buffer TE or nuclease-free water to the center of the membrane (Pre-heat the Buffer TE or nuclease-free water at 65℃ can improve elution efficiency).

15. Centrifuge at 12,000 rpm for 2 minutes to collect DNA. If more yield is needed, the flow-through can be re-added into the center of the membrane and let it stand for 5 minutes at room temperature and centrifuge at 12,000 rpm for 2 minutes to elute the DNA again.

Note:

1. Please read the Product Manual carefully before use.

2. Fresh materials should be prepared to ensure that the genomic DNA is not degraded.

3. Repeated freezing and thawing of samples should be avoided, as this may result in a decrease in the yield of DNA.

4. Do not use too much bacterial is better for fully lyse, otherwise it may affect the yield and purity of genomic DNA.

5. For your safety and health, please wear safety glasses, gloves, or protective clothing.

For Research Use Only!