MagBind Blood/Tissue/Cell Total DNA Kit

 

Product Name	Cat. No.	Spec.
MagBind Blood/Cell/Tissue Total DNA Kit	G3604-50T	50 T

 

 

This kit is a broad-spectrum kit for the purification ofDNA from animal/plant tissues,fresh, whole mammalian blood and cultured cells. The kit provides a specially optimized lysis buffer torelease genomic DNA, superparamagnetic magnetic beadstoselectively adsorb genomic DNA in thebinding buffer, washing buffer toremove impurities, andeluent to elute adsorbed genomic DNA. High-purity intact genomic DNA can be rapidly extracted from 2-30 mg of animal tissues, 106-107 fresh cultured cell suspension, and 5-100 μL of whole blood ofanucleated or nucleated red blood cells (including anticoagulants). Thepurified genomic DNA can be directly applied in PCR and Real-time PCR analysis,enzyme digestion,SNP genotype analysis andother molecular biology experiments.

 

 

Proteinase K andRNaseA are shipped with wet ice, stored at -20℃; Other reagents are shipped andstored at room temperature; validfor up to 1 year.

 

 

Component Number	Component	G3604-50T
G3604-1	Buffer GA	12mL
G3604-2	Proteinase K	1mL
G3604-3	RNase A	200 μL
G3604-4	Buffer GB	12mL
G3604-5	SweMag Beads	1 mL
G3604-6	Buffer PD	12 mL
G3604-7	Buffer PW	24mL
G3604-8	Buffer TE	10mL
Manual		One copy

 

 

1. Ifa precipitate has formed in BufferGA, warm it at65°C until the precipitate has fully dissolved.

2. Before first use, add18 mL of absolute ethanol to Buffer PD,add56 mL of absolute ethanol to Buffer PWandmix well separately.

3. Magnetic stands and absolute ethanol are required but not supplied in this kit.

 

 

1. Samplespreparation:

a. Tissue sample homogenate:

Note: The amount of tissue should not exceed30 mg, otherwise the yield and quality ofDNA may decrease.

Take out fresh or frozen tissue at -80°C, placethemin a 1.5 mL centrifuge tube or specialgrindingtube containing 2-3 grinding beads with a diameter of 3 mm(G0203-150G recommended), add 200 μL of Buffer GA, and use aTissue 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. Cell sample treatment:

Take cell suspension of cell density at106-107, place in a 1.5 mL centrifuge tube, centrifuge at 1,000 rpm for 5 min,and carefully remove all the supernatant of the culture medium. Then, add 200 μL of Buffer GA to the centrifuge tube and mix by vortexing;

c. Whole blood of anucleated red blood cells treatment:

Take 50-100 μL of whole blood ofanucleated red cells (including anticoagulant) in a 1.5 mL centrifuge tube, add Buffer GA to 200 μL, mix by vortexing. (if processing a larger volume of blood, add 3 volumes of red blood celllysis buffer (G2015 recommended) to the sample and mix upside down, place at room temperature for 5 min, mix inverted 2-3 times duringincubation, centrifuge at 3,000 rpm for 10 min, gently aspirate the supernatant using a pipette, do not aspirate the leukocyte pellet, add 200 μL Buffer GA,pipet thelysate up and down several timestomix well.

d. Whole blood of nucleated red blood cells treatment:

Take 5-20 μLofwhole blood of nucleated red blood cell (containing anticoagulant), place it in a 1.5 mL centrifuge tube, add Buffer GA to 200 μL, and mix well by pipetting;

2. Add20 μL Proteinase K and 4μL of RNase A to the sample from step 1, incubate at56℃ for 30 minutes (Mix every 10 minutes to accelerate tissue lysis).

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

3. Centrifuge at 12,000 rpm for2 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, andincubate at 70 °C for 10 min.

5. Add200 μL of absolute ethanol to thecentrifuge tube, mix upside and down(any visible precipitate that may form after addingisopropanol).

6. Centrifuge at 12,000 rpm for 2 min and transfer the supernatant to a Nuclease-free centrifuge tube, avoiding aspirating the pellet.

7. add 20 μL SweMag Beads to the mixture (SweMag Beads must befully resuspended andevenly distributedprior to use), and mix well with pipette.

8. Stand atroom temperaturefor 10 minutes and mix well with pipette every 2-3 minutes to help SweMag Beadsevenly distributed.

9. Place the centrifuge tube on a magnetic stand for30 seconds. When the magnetic beads are completely attached, carefullydiscard the supernatant with a pipette (To avoid lower recovery rate, do not aspirate the magnetic beads).

10. Add500 µLofBufferPD, removemagnetic stand andpipet up and down gently to resuspend the magnetic beads. And then place the tube on a magnetic stand for 30 seconds.When the magnetic beads are completely attached, carefullydiscard the supernatant with a pipette.

11. Add600 µLofBufferPW, removemagnetic stand andpipet up and down gently to resuspend the magnetic beads. And then place the tube on a magnetic stand for 30 seconds.When the magnetic beads are completely attached, carefullydiscard the supernatant with a pipette.

12. Repeat step 11.

13. Open the centrifuge tube lid, leave it at room temperature for 5-10 minutes to ensure complete evaporation of residual ethanol and other liquids. (Do not allow them to dry out as this renders them non-functional.)

14. Remove the magnetic stand, add 50–100 μL of Nuclease-free Water orBuffer TEand pipet up and down gently to resuspend the magnetic beads, stand at room temperature for 5 minutes.

15. Place the tube on a magnetic stand to collect the beads against the side of the tube, then aspirate the supernatant to a new centrifuge tube. The supernatant contains the purified DNA.

 

 

1. Please readtheProduct Manual carefully before use.

2. Fresh materials should bepreparedto ensure that the genomic DNA is not degraded.

3. Do not freeze the beads as this irreparably damages them.

4. Samples should avoid repeated freeze-thaw cycles, as this will result in a decrease DNA yield

5. Always keep the beads in solution. Do not allow them to dry out as this renders them non-functional.

6. When using the beads, resuspend thoroughly in the storage buffer by vortexing before removal.

7. For your safety and health, pleasewearsafety glasses, gloves, or protective clothing.

 

For Research Use Only!