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Code: CSB-PPM02005
Sample: up to 200 mL bacterial cells
Yield: up to 1.5 mg of plasmid
As research moves from DNA sequencing to analyzing of gene function, the needs of isolating large quantities of high-quality plasmid DNA by rapid methods has been increasing.
CUSABIO Plasmid DNA Purification Maxiprep Kit is designed to isolate high-quality plasmid DNA by our unique plasmid purification beads and combine with traditional isopropanol precipitation method, which also incorporates a unique Endotoxin Removal Wash designed to remove substantial amounts of protein, RNA and endotoxin contaminants from purified plasmid DNA.
The extracted plasmid DNA is suitable for various applications, including restriction enzyme digestion, PCR, sequencing, ligation, as well as transformation and transfection in a variety of cell experiments.
Plasmids extracted with the CUSABIO Plasmid DNA Purification Maxiprep Kit at a supersampling volume of 1 ug.
| Each Component | 5T |
|---|---|
| Buffer S1 | 60 mL |
| Buffer S2 | 60 mL |
| Buffer S3 | 60 mL |
| RNase A | 18 mg |
| Wash Buffer | 100 mL*2 |
| ER Buffer | 18 mL |
| Elution Buffer | 60 mL |
| TE Buffer | 5 mL |
| Plasmid Purification MaxiPrep Column | 5 |
RNase: Store at -20°C upon receipt.
All other components: Store at 15-25°C upon receipt.
Q: What if the endotoxin residue in the extracted plasmid is too high?
A: This kit has a unique Endotoxin Removal Wash, which effectively removes endotoxin so that the plasmid DNA obtained can be of cellular grade.
Q: During plasmid lysis, what is the normal state of the solution S2 after addition?
A: After the addition of S2, the solution becomes light yellow and clarified, with a slightly viscous texture, and slightly stretched when the cap is opened.
Q: During the process of plasmid neutralization, what is the normal state of solution S3 after addition?
A: After S3 is added, the solution shows white eggflower-like precipitation, at this time, we need to observe whether there is still a viscous mass after P2 solution is added, if there is, we need to add a large amount of mixing evenly.
Q: Why does S2 solution precipitate white flocculent in fall and winter?
A: S2 solution is composed of SDS and NaOH, of which SDS is easy to crystallize and precipitate at low temperatures. Before use, it needs to be put into a 37℃ oven or water bath 30-60min beforehand until the reagent is clarified and transparent again.
Q: What should I do if the plasmid supernatant is still slightly turbid after centrifugation?
A: If the problem occurs in the batch, you can consider whether the centrifugation time is too short, you can increase the centrifugation time to see if it is too short; if it is a turbid sample, you can re-centrifuge the sample into EP tubes for 3-5min, and then suck up the supernatant for filtration; if the supernatant of individual samples is clarified to the naked eye, but it is very clogged during filtration, you can centrifuge the sample again and then re-filter the supernatant.
Q: What should I do if there is RNA residue when running the plasmid?
A: If there is RNA residue in the batch, you can reduce the amount of isopropanol appropriately, under normal circumstances, the amount of isopropanol is 70% of the eluent, but too much isopropanol will adsorb too much DNA and RNA fragments, and the specific amount can be adjusted according to the specific experimental situation; In addition, there is also the possibility that the RNA enzyme is inactivated, and the RNA is not digested completely when the bacterium is resuspended in the first step.
CUSABIO Plasmid DNA Purification Maxiprep Kit Protocol
