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.
Key Features and Advantages
Unique Technological Design: Combines unique plasmid purification beads with the traditional isopropanol precipitation method for ensuring high-quality plasmid DNA purification.
Lower Endotoxin: Unique endotoxin removal wash to make sure effectively removes substantial amounts of protein, RNA, and endotoxin contaminants, enhancing the purity of the isolated plasmid DNA.
High DNA Yield: Capable of purifying up to 1.5mg of plasmid DNA from a 200ml overnight bacterial culture transformed with a high-copy-number plasmid.
The yield of plasmid DNA from the same sample: CUSABIO > A Company > B Company > C Company > D Company.
The endotoxin level validation results in ascending order are: CUSABIO < A Company < D Company < B Company < C Company.
Applications
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.
Product Details
General Steps of Operation
Experimental Operation Video
Using the CUSABIO Plasmid Maxiprep Kit: A Step-by-Step Guide
Supporting Data and Figures
Plasmids extracted with the CUSABIO Plasmid DNA Purification Maxiprep Kit at a supersampling volume of 1 ug.
Product Components
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
Storage Conditions and Product Stability
RNase: Store at -20°C upon receipt.
All other components: Store at 15-25°C upon receipt.
FAQ
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.
Protocol
CUSABIO Plasmid DNA Purification Maxiprep Kit Protocol
