Magnetic Bead-based DNA/RNA Extraction Kit, 50 T (for Nucleic Acid Extraction)

Product Information

Description/Introduction

The kit is designed for rapid isolation of various viral DNA/RNA from serum, plasma, urine, supernatant of cell culture medium, virus stock and infected tissues. Through the superparamagnetic beads independently developed by our company, under the action of a special buffer system, nucleic acid is specifically adsorbed on the surface of magnetic particles, and proteins, cell fragments and other pollutants can be effectively rinsed. Finally, high quality DNA/RNA is obtained by Nuclease-free Water elution. The obtained high quality DNA/RNA can be directly used in downstream PCR, cDNA synthesis, RT-qPCR and other molecular biology experiments.

Storage and Handling Conditions

Proteinase K and Carrier RNA are shipped with wet ice and stored at -20°C. Other reagents are shipped and stored at room temperature; valid for 12 months.

Product Contents

Before starting (please read carefully)

1. If Buffer MVL and Buffer MPA precipitates, please heat it at 37°C and use it when it is restored to room temperature.

2. Please add 18 mL of anhydrous ethanol to Buffer MPA and 60 mL anhydrous ethanol to Buffer MPB before first use.

3. Carrier RNA can be sub-packed and stored at -20°C before first use, and repeated freezing and thawing should be avoided.

4. Please pre-cool the grinder if grinding tissue with a grinder.

5. Magnetic stands are required but not supplied in this kit.

Assay Protocol / Procedures

1. Samples preparation:

a. Lysis of serum, plasma, urine, supernatant of cell culture medium and virus stock: Sampling 10~200 μL of serum, plasma, urine, supernatant of cell culture or virus stock. The initial amount of less than 200 μL can be replenished to 200 μL with PBS or Nuclease-free Water.

b. Lysis of infected tissue: Sampling 10~20 mg fresh or cryopreserved infected tissues and move them in 1.5 mL Nuclease-free centrifuge tubes containing 2~3 pcs 3 mm grinding beads (recommended G0203) or use special grinding tubes (recommended HT-200-M). Immediately place the centrifuge tube or grinding tube containing the tissue in liquid nitrogen, and then grind the tissue thoroughly to homogenate at low temperature using a tissue grinder (recommended KZ-5F-3D, if the tissue is not thoroughly homogenized, the yield and quality of DNA/RNA will be affected). Add 200 μL of PBS or Nuclease-free Water after grinding.

2. Add 200 μL of Buffer MVL, 20 μL of Proteinase K and 1 μL of Carrier RNA. After mix thoroughly, incubate at 56°C for 10 min.

3. Add 200 μL of anhydrous ethanol, mix upside down, and then add 20 μL of SweMag Beads (SweMag Beads should be vortexed until evenly dispersed before use), upside down several times until the beads are uniformly dispersed.

4. Stand at room temperature for 10 min, and mix 2~3 times with a pipette or vortex mixer until the beads are uniformly dispersed.

5. Move the centrifuge tube to the magnetic stand and let stand for 30 s until the magnetic beads were fully adsorbed, and reverse the centrifuge tube gently up and down several times with the magnetic stand to wash down the magnetic beads left on the pipe wall. When it is clear, absorb and discard the supernatant (in order to avoid affecting the extraction efficiency, do not suck the magnetic beads out).

6. Remove the magnetic stand, add 500 μL of Buffer MPA, gently blow with a pipette until the magnetic beads are evenly dispersed, then move the centrifuge tube to the magnetic stand and let stand for 30 s, and reverse the centrifuge tube gently up and down several times with the magnetic stand to wash down the magnetic beads left on the pipe wall. After it is clear, discard the supernatant (in order to avoid affecting the extraction efficiency, do not suck the magnetic beads out).

7. Remove the magnetic stand, add 700 μL of Buffer MPB, blow it with a pipette until the magnetic beads are dispersed evenly, then move the centrifuge tube to the magnetic stand for 30 s, and reverse the centrifuge tube gently up and down several times with the magnetic stand to wash down the magnetic beads left on the pipe wall, after it is clear, discard the supernatant (in order to avoid affecting the extraction efficiency, do not suck the magnetic beads out).

8. Repeat step 7.

9. Open the centrifuge tube lid, leave it at room temperature for 5~10 minutes to make the ethanol evaporate completely.

10. Remove the magnetic stand, add 50~80 μL of Nuclease-free Water to the centrifuge tube, gently blow until the magnetic beads are uniformly dispersed with a pipette, stand at room temperature for 3 min.

11. Move the centrifuge tube to the magnetic stand until the magnetic beads are completely adsorbed, and absorb the supernatant into a new nuclease-free centrifuge tube to obtain high purity DNA/RNA.

Note

1. Please read the Product Manual carefully before use.

2. Due to the addition of Carrier RNA in the extraction process, the quantity can not be determined by electrophoresis or absorptometer.

3. The Carrier RNA provided by this kit is the RNA from E. coli, which is mainly to improve the recovery efficiency of trace nucleic acid and prevent the degradation of trace RNA. If the PCR primers have high homology with Carrier RNA, the primers can be redesigned.

4. Magnetic beads are precipitate easily, and should be vortexed until they are well dispersed before use.

5. Magnetic bead suspension should avoid freezing during preservation.

6. Before adding magnetic beads to the sample, it is necessary to mix the sample with other reagents evenly.

7. Before elution, ethanol should be completely volatilized to avoid the influence of residual ethanol on downstream experiments.

8. For your safety and health, please wear a lab coat and disposable gloves.

For Research Use Only!

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