Universal DNA Purification and Gel Extraction Kit with Magnetic Beads, 10 T (for Nucleic Acid Extraction)

Product Information

Description/Introduction

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.

Storage and Handling Conditions

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

Product Contents

Before starting (please read carefully)

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

2. Add all the RNase A provided in the kit to Buffer SP1 before use, and it can be stored at 4°C for 6 months.

3. After using SP2, the lid should be closed immediately to avoid long-term contact with air.

4. Please pre-cool SP3 at 4°C before use.

5. Please add 77 mL anhydrous ethanol to Buffer PW before use.

Assay Protocol / Procedures

1. Harvest bacterial culture from 100 mL (200 mL of overnight cultured bacterial solution is recommended for low copy plasmid) of overnight cultured bacterial solution by centrifugation at 10,000 rpm (11,500 ×g) for 3 min at room temperature. Collect the culture in the 50 mL centrifuge tube (remove the supernatant as much as possible).

2. Add the 8 mL of SP1 (please check whether or not to add the RNase A first), use a pipette or vortex oscillator to thoroughly suspend the bacteria (be sure to disperse the bacteria thoroughly, otherwise it will affect the lysis, resulting in low quality and purity of the extracted plasmid).

3. Add 8 mL of SP2, immediately and gently turn it upside down for 8-10 times to make the bacteria fully lysed (this step should not be violently shaken, and should be done within 5 min). At this point, the solution should become clear and sticky, if it does not become clear, it may be too many bacteria, then reverse it a few times until the solution is transparent.

4. Add 8 mL of SP3 (pre-cool at 4°C in advance), immediately and gently upside down 10~12 times, the solution appears compact agglomerate, place 5 min on the ice, centrifuge at 10,000 rpm for 20 min at 4°C, and take the supernatant in a Nuclease-free 50 mL centrifuge tube (self-provided).

5. Add 10 mL of SPD and 15 mL of anhydrous ethanol, upside down and mix thoroughly. Then add 600 μL of SweMag Beads (Disperse the magnetic beads evenly by vortex before use), upside down until the beads are uniformly dispersed, place the SPD at room temperature for 30 min and mix upside down every 3~5 min during the period.

6. After placing the 50 mL centrifuge tube on a magnetic stand for 2 minutes，turn the centrifuge tube up and down gently with the magnetic stand several times to rinse the magnetic beads remaining on the pipe cover, continue to stand until the liquid clarifies and discard the liquid.

7. Remove the centrifuge tube from the magnetic stand, add 5 mL of PW (check whether anhydrous ethanol is added), gently upside down to make the magnetic beads disperse evenly, then place the centrifuge tube on the magnetic stand, gently reverse the centrifuge tube up and down several times with the magnetic stand, rinse the magnetic beads remaining on the pipe cover, continue to rest until the liquid is clarified and discard the liquid.

8. Repeat step 7.

9. Open the centrifuge tube lid, place 5~10 min at room temperature or 3~5 min at 65°C to make the ethanol evaporate completely (avoid excessive drying of magnetic beads, so as not to affect the yield of nucleic acid).

10. Remove the magnetic stand, add 1~3 mL Buffer TE or Nuclease-free Water, gently blow with the pipette until the beads are evenly dispersed, and place at room temperature for 3~5 min.

11. Reposition the centrifuge tube on the magnetic stand until the magnetic beads are completely adsorbed, the supernatant was taken in a new Nuclease-free centrifuge tube to obtain high concentration and high purity plasmid DNA.

12. Plasmid DNA is stored at -20°C for backup.

Note

1. Please read the Product Manual carefully before use.

2. Magnetic bead suspension should avoid freezing during preservation.

3. Magnetic beads are precipitate easily, so vortex thoroughly before use.

4. Ethanol should be completely volatilized before eluting the plasmid to avoid the influence of residual ethanol on downstream experiments.

5. Please do not dry the magnetic beads for a long time, so as not to affect the elution efficiency of DNA.

6. If the plasmid DNA is to be preserved for a long time, it is recommended to use Buffer TE elution.

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

For Research Use Only!

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