SKU : G3600-100T
Categories : Biochemicals , 1. Chemical and Reagents , Buffers , Servicebio ,
Brand : Servicebio
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Product Information
Product Name | Cat. No. | Spec. |
MagBind Plasmid DNA Miniprep Kit | G3600-100T | 100T |
Product Description/Introduction
MagBind Plasmid DNA Miniprep Kit employs unique magnetic beads based on modified alkali lysis method to perform stable, efficient and convenient extraction of small amounts of plasmids from E. coli. The DNA yield is up to 20 μg extracting from 1-5mL fresh bacterial culture.
Plasmids extracted by this kit can be directly used for cell transfection, DNA sequencing, PCR, PCR-based mutagenesis, in vitro transcription, bacterial transformation, endonuclease digestion and other experiments.
Storage and Shipping Conditions
RNase A should be shipped with wet ice, and stored at -20; Other reagents are shipped and stored at room temperature for up to 1 year.
Product Contents
Component Number | Component | G3600-100T |
G3600-1 | Solution I | 20 mL |
G3600-2 | Solution II | 20 mL |
G3600-3 | Neutralization Buffer | 20 mL |
G3600-4 | RNase A | 200 μL |
G3600-5 | SweMag Beads | 3.5 mL |
G3600-6 | Binding Buffer | 20 mL |
G3600-7 | Buffer SPW | 24 mL |
G3600-8 | Buffer TE | 15 mL |
Manual | One copy | |
Before starting (please read carefully)
1. Before first use, add all of the RNase A provided in the kit to Solution I, mix well, and mark the vial. Store at 4 after adding RNase A.
2. At low temperatures, Solution II may precipitate. In this case, redissolve the precipitates at 65 and mix well prior to use.
3. Do not shake Solution II (Lysis Buffer) vigorously, as it may produce large amounts of air bubbles. After use, tightly cap the bottle to prevent acidification by CO2 in the air.
4. Before first use, add the specified amount of absolute ethanol to Binding Buffer and Buffer SPW,mix well, and mark the bottle.
5. Magnetic stands, absolute ethanol and sterilized 1.5 mL centrifuge tubes are required but not supplied in this kit.
Assay Protocol / Procedures
1. Centrifuge 1-5 mL of overnight fresh bacterial culture at 12,000 rpm for 1 minute and discard the supernatant.
2. Resuspend the bacterial pellet thoroughly in 150 µL of Solution I (please ensure RNase A has been added before using) by vortexing or pipetting up and down.
3. Add 150 µL of Solution II and mix gently by inverting the tube 8-10 times to ensure complete lyse of bacteria.
Note 1: Mix well gently, do not shake violently to avoid genomic DNA breakage and contamination of the extracted plasmid.
Note 2: Afetr 8-10 times inversions, the mix shouled be clear. And the total lysis time should not exceed 5 minutes.
4. Add 150 µL of precooled Neutralization Buffer and gently invert the centrifuge tube 8-10 times to mix well immediately (A small amount of tiny white precipitate appear). Centrifuge at room temperature at 12,000 rpm for 10 minutes.
5. Transfer carefully the supernatant to a new 1.5 mL centrifuge tube, add 450 uL of Binding Buffer and mix it upside and down. And then add 30 μL of SweMag Beads suspension (SweMag Beads must be fully resuspended and evenly distributed prior to use), gently invert up and down several times, until the magnetic beads distribute evenly.
6. Stand at room temperature for 5 minutes. Mix 2-3 times with pipette or vortex mixer during incubation, to distribute evenly the magnetic beads.
7. Place the tube on a magnetic stand and stand for 30 seconds until the magnetic beads are completely attached. Gently invert the magnetic stand with the centrifuge tube up and down several time to wash away residual magnetic beads. Aspirate and discard the clear supernatant (To avoid lower recovery rate, do not aspirate the magnetic beads).
8. Add 300 μL Buffer SPW, remove the tube containing the pelleted magnetic beads from the magnetic stand, pipet up and down gently to resuspend the magnetic beads. Place the tube on a magnetic stand and stand for 30 seconds, Gently invert the magnetic stand with the centrifuge tube up and down several times to to wash away residual magnetic beads and salt. Aspirate and discard the clear supernatant (To avoid lower recovery rate, do not aspirate the magnetic beads).
9. Repeat step 8.
10. Open the centrifuge tube lid, place it at 65 for 3-5 minutes or 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.)
11. Remove the magnetic stand, add 5080 μL Buffer TE or Nuclease-free Water and pipet up and down gently to resuspend the magnetic beads, stand at room temperature for 3 minutes.
12. Place the tube on a magnetic stand until the magnetic beads are completely attached, then aspirate the supernatant to a new centrifuge tube. The supernatant contains the purified plasmids.
Note
1. Please read the Product Manual carefully before use.
2. Do not freeze the beads as this irreparably damages them.
3. Always keep the beads in solution. Do not allow them to dry out as this renders them non-functional.
4. When using the beads, resuspend thoroughly in the storage buffer by vortexing before removal.
5. Before adding the magnetic beads, other reagents need to be mixed well.
6. Ethanol should be completely evaporate before plasmid DNA elution due to residual ethanol affecting downstream experiments.
7. For your safty and health,please wear safety glasses, gloves, or protective clothing.
For Research Use Only!
