MagBind Plasmid DNA Miniprep Kit

 

Product Name	Cat. No.	Spec.
MagBind Plasmid DNA Miniprep Kit	G3600-100T	100T

 

 

MagBind Plasmid DNA Miniprep Kit employsunique magnetic beadsbased 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 20μgextractingfrom 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.

 

 

RNase A should be shipped withwet ice, and stored at-20℃; Other reagents are shipped and stored at room temperaturefor up to 1 year.

 

 

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

 

 

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 toBinding Buffer andBuffer 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.

 

 

1. Centrifuge 1-5 mL of overnightfreshbacterial cultureat 12,000 rpm for 1 minute and discard the supernatant.

2. Resuspend thebacterial pelletthoroughlyin150 µL ofSolution I (please ensure RNase A has been added before using) by vortexing or pipetting up and down.

3. Add150 µL ofSolution II and mix gently by inverting the tube8-10 times to ensurecomplete lyseofbacteria.

Note 1:Mix well gently, do not shake violently to avoid genomic DNA breakage and contamination of the extracted plasmid.

Note 2:Afetr8-10 times inversions, the mix shouled be clear. And the total lysis time should not exceed 5 minutes.

4. Add150 µL ofprecooledNeutralization Buffer andgently invert the centrifuge tube 8-10times to mix well immediately (A small amount of tiny white precipitate appear).Centrifuge at room temperature at 12,000rpm for10 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 add30 μL ofSweMag Beads suspension (SweMag Beads must befully resuspended and evenly distributedprior to use), gentlyinvert up and down several times, until the magnetic beadsdistribute evenly.

6. Stand at room temperature for 5 minutes. Mix 2-3 times with pipette or vortex mixer during incubation,todistribute evenly the magnetic beads.

7. Place the tube on amagnetic stand and stand for 30 secondsuntil the magnetic beads are completelyattached. Gently invertthe magneticstand with the centrifuge tube up and down several time towash awayresidual magnetic beads.Aspirate and discard theclearsupernatant (To avoid lower recovery rate, do not aspirate the magnetic beads).

8. Add300 μL Buffer SPW, remove the tube containing the pelleted magnetic beads from themagnetic stand,pipet up and down gently to resuspend the magnetic beads. Place the tube on amagnetic stand and stand for 30 seconds, Gently invertthe magneticstand with the centrifuge tube up and down several timestotowash awayresidual magnetic beads and salt.Aspirate and discard theclearsupernatant (To avoid lower recovery rate, do not aspirate the magnetic beads).

9. Repeat step 8.

10. Open the centrifuge tube lid, placeitat65℃ for3-5 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 themagneticstand, add50–80 μL Buffer TE orNuclease-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 standuntil the magnetic beads are completelyattached, then aspirate the supernatant to a new centrifuge tube. The supernatant contains the purified plasmids.

 

 

1. Please readtheProduct 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 completelyevaporate before plasmid DNA elution due to residual ethanol affecting downstream experiments.

7. For your safty and health,pleasewearsafety glasses, gloves, or protective clothing.

 

For Research Use Only!