MagBind HighPure Plasmid DNA Maxiprep Kit

 

Product Name	Cat.No.	Spec.
MagBind HighPure Plasmid DNA Maxiprep kit	G3610-10T	10T

 

 

Making use of the reversibly binding nucleic acid characteristics of supercis magnetic beads and its unique buffer system, the kit can quickly and efficiently obtain 600~1,500 μg high purity plasmid DNA from the overnight cultured bacterial solution of 100 mL. The plasmid extracted by this kit can be used in molecular biology experiments such as restriction enzyme digestion reaction, ligation reaction, PCR amplification, sequencing and so on.

 

 

RNase A is shipped with wet ice and stored at -20℃. Other reagents are shipped and stored at room temperature; valid for 12 months.

 

 

Component Number	Component	G3610-10T
G3610-1	Buffer SP1	90 mL
G3610-2	Buffer SP2	90 mL
G3610-3	Buffer SP3	90 mL
G3610-4	RNase A	180 μL
G3610-5	Buffer SPD	110 mL
G3610-6	Buffer PW	33 mL (77 mL of anhydrous ethanol added before use)
G3610-7	Buffer TE	30 mL
G3610-8	SweMag Beads	7 mL
Manual		One copy

 

 

1. Prepare heating blocks at 80°C and 55°C in advance.

2. If Buffer CRL and Buffer FRB precipitate, please heat at 65°C to dissolve and use it after returning to room temperature.

3. Prepare only part of the DNase working solution required for the experiment.

4. Please add 64 mL anhydrous ethanol to Buffer RW2 before first use, mix it thoroughly and then use it.

 

 

1. Samples preparation:

a. Paraffin sections: scrape 5~8 paraffin sections (5~10 μm thick, 1×1 cm2 size). If the sample is exposed to air, the 2~3 pieces on the surface will be discarded.

b. Paraffin blocks: scrape the tissue sample of about 30 mg with a sterilized scalpel, remove the excess paraffin as much as possible and cut up the sample.

c. Sample tissue soaked in formalin: dry the liquid on the sample surface with filter paper, take about 30 mg sample, cut it up and place it in 1.5 mL centrifuge tube, add 500 μL 1×PBS buffer (pH 7.4), vortex oscillation for 10 s, then centrifuge at 12,000 rpm for 1 min at room temperature, discard the supernatant, repeat 3 times.

2. Transfer the sample to a 1.5 mL centrifuge tube, add 500 μL Buffer DP, incubate at 80°C for 3 min, and vortex oscillation for 10 s while the sample is hot.

3. Add 200 μL Buffer CRL to the centrifuge tube, mix thoroughly with vortex, centrifuge at 12,000 rpm for 1 min at room temperature, and form two layers of solution (upper layer is oil phase, lower layer is water phase).

4. Add 20 μL Proteinase K to the lower aqueous phase, gently mix with a pipette (try not to destroy the stratification), incubate at 55°C for 15 min.

5. Then transfer the centrifuge tube to a 80°C heating block, incubate for 15 min (if there is only one heating block, put the centrifuge tube containing the sample at room temperature first, and then put the centrifuge tube into the heating block when the temperature reaches 80°C), mix it upside down.

6. Centrifuge at 12,000 rpm for 5 min at room temperature, and form two layers of solution (upper layer is oil phase, lower layer is water phase).

7. Remove the aqueous solution from the lower layer into a new Nuclease-free 1.5 mL centrifuge tube (do not get the upper oil phase solution and other impurities). Add Buffer FRB with equal volume of aqueous phase and anhydrous ethanol with 3 times volume of aqueous phase (there may be precipitation), and use liquid pipette to blow and mix well.

8. Transfer all the mixture to RNA Spin Column, centrifuge at 12,000 rpm for 2 min at room temperature, and discard the filtrate.

9. Prepare DNase working solution: take 35 μL Nuclease-free Water, 5 μL 10×DNase Reaction Buffer and 10 μL DNase in a new Nuclease-free centrifuge tube, gently mix with a pipette.

10. Add the DNase working solution droplet to the center of the membrane of the RNA Spin Column and stand at room temperature for 15 minutes.

11. Add 500 μL Buffer RW1 to RNA Spin Column, centrifuge at 12,000 rpm for 1 min at room temperature, and discard the filtrate.

12. Add 600 μL Buffer RW2 to RNA Spin Column (please add Buffer RW2 along the pipe wall to help flush the residual salt on the pipe wall), centrifuge at 12,000 rpm for 1 min at room temperature, and discard the filtrate.

13. Repeat step 12.

14. Place RNA Spin Column on the collection tube and centrifuge at 12,000 rpm for 2 min at room temperature. Take out the RNA Spin Column and put it on a new Nuclease-free 1.5 mL centrifuge tube.

15. Open the lid of RNA Spin Column, leave it at room temperature for 3~5 minutes to make the ethanol evaporate completely.

16. Add 50~100 μL of Nuclease-free Water to the center of the membrane of the RNA Spin Column, stand at room temperature for 5 minutes, centrifuge at 12,000 rpm for 2 min to collect RNA solution. To get a higher concentration of RNA, you can also add the first eluent back to RNA Spin Column, then stand at room temperature for 5 min and centrifuge for 2 min to elute RNA again.

 

 

1. Please read the Product Manual carefully before use.

2. The yield and integrity of RNA extracted by this kit depend on the type of sample, fixed time, fixed condition and sample storage time. The fixed time is preferably within 8~24 hours. If the fixed time of the sample is more than 24 hours or the storage time is more than 1 year, it may lead to a large number of RNA degradation.

3. The samples should be completely dehydrated before embedding to prevent the residual formalin from affecting the follow-up experiments.

4. When sampling embedded tissue, we should remove the excess paraffin and cut up the sample as much as possible, and the sampling amount should not exceed 30 mg, otherwise it is easy to cause insufficient lysis and affect the yield of nucleic acid.

5. Nucleic acids purified from FFPE samples are not recommended for downstream applications that require full-length RNA.

6. Wear lab clothes, disposable gloves and masks during the experiment, avoid talking, and use Nuclease-free pipette tips and centrifuge tubes to avoid cross contamination.

7. RNA extraction special operation test bench and electrophoresis equipment should be used.

 

For Research Use Only!