Plasmid DNA extraction and purification magnetic beads kit
This kit, with proprietary separation and buffer systems, allows extracting and purifying plasmid DNA from 2.0~5.0 mL of bacteria culture. The typical yield is 10~20 µg of high quality plasmid DNA, and the typical OD260/280 is between 1.75 and 1.85.
Purified plasmid DNA can be directly used for a variety of molecular biology applications such as enzymatic digestion, sequencing and transformation.
The kit will work with a 48 well round bottom plates if a special magnetic frame is used.
The kit can also be used with a variety of automatic nucleic acid extraction instruments or workstations. |
Catalog # |
Si-Mag-PL1 |
Precautions |
- Wear protective gloves, clothing, eye, and face protection. Wash hands thoroughly after handling.
- Avoid freeze/thaw cycles and centrifugation which could damage the beads.
- Vortex samples for about 10 seconds before adding magnetic beads.
- Vortex beads and mix them well with DNA containing material to ensure best performance.
- Elute DNA from the beads completely.
|
Kit Components included |
- Si-Mag magnetic beads 1.5 mL
- RNase A solution 60 uL
- Suspension solution 20 mL
- Lysis solution 20 mL
- Neutralization solution 35 mL
- Wash solution 25 mL (Add 25 ml of Isopropanol before use)
- Elution Buffer 10 mL
|
Materials needed but not provided with the kit |
- 80% Ethanol in water.
- Si-Mag Magnet (sold separately) or other magnetic racks compatible with vials used.
- Isopropanol (ACS grade).
|
Storage |
Magnetic beads should be stored at 2-8°C but other kit reagents need to be stored at room temperature. Lysis solution may turn cloudy if store in th cold room and to clear it up place the bottle into a water bath at 37°C. |
Protocol |
- Preparation. Before the first use, add all the RNase A solution into the Suspension Solution.
- Harvest cells. Spin 2-5 mL of overnight grown bacterial cells at 12,000 rpm for 2 min.
- Re-suspend the cells. Remove culture media completely. Add 200 uL of suspension solution (with RNase A), then vortex to ensure complete suspension of cell. Transfer cells to a clean Eppendorf tube.
- Lyse the cells. Add 200 uL of Lysis solution and mix by inverting the tube for 6-8 times, then incubate for 2 min. Do NOT vortex.
- Neutralize the solution. Add 350 uL of Neutralization solution and mix by inverting the tube for 8-10 times. Do NOT vortex.
- Spin the tube at 12,000 rpm for 15 min at room temperature.
- Transfer the solution to a clean Eppendorf tube, then add 15 uL of magnetic beads, mix well and incubate 3-5 min at RT. Put Eppendorf tube onto the Si-Mag magnet rack for 20 seconds.
- Remove solution by holding the magnet rack upside down or by pipetting.
- Wash the beads with 500 uL of Wash Solution and then repeat
Step 8.
- Wash the beads with 500 uL of 80% ethanol twice repeating
Step 8.
- Dry the beads at 55°C for 8 min leaving the tube open. Do not over-dry
the beads.
- Elute the DNA from beads with 50-100 uL of elution buffer, incubate for at least 2 min and then vortex at full speed for 1 min. Alternatively, incubation at 60°C for 2 min may improve the recovery for DNA larger than 10 kb.
- Remove beads by using magnet rack, pipette DNA out and transfer to a clean tube.
- Store purified DNA at -20°C for long-term storage.
|
| |
