CAR-Manu™ High Pure Plasmid Isolation Kit (CAR-K-Z021)

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Product Name
CAR-Manu™ High Pure Plasmid Isolation Kit

Cat. No.
CAR-K-Z021

General Description
Low to medium throughput, mini scale, plasmid isolation.

Product Summary
The high-purity plasmid isolation kit uses alkaline lysis to isolate a small amount of purified plasmid DNA. This method is a common method for producing high-purity plasmid DNA from E. coli and is not subject to RNA contamination.

In the presence of chaotropic salt (guanidine hydrochloride), nucleic acids bind to the surface of the glass fiber fleece. This allows high-purity filter tubes to specifically fix nucleic acids (DNA and RNA) while removing contaminants.

Components
• Suspension Buffer
• RNase A, dry powder
• Lysis Buffer
• Binding Buffer
• Wash Buffer I
• Wash Buffer II
• Elution Buffer
• High Pure Spin Filter Tubes (containing glass fiber fleece)
• Collection Tubes

Capacity
The high-purity spin filter tube can hold a sample volume of 700 μL

Applications
The high-purity plasmid isolation kit can prepare up to 15 μg of purified plasmid DNA from bacterial cultures and can be used directly in most molecular biology applications:
•PCR
•Sequencing
•clone
• In vitro transcription
• Restriction enzyme digestion
• Randomly infused labels

Preparation Note
The kit relies on alkaline lysis to release plasmid DNA from bacteria. RNase removes all RNA from the lysate. After removing cell debris and (captured) genomic DNA by centrifugation, the remaining supernatant is mixed with chaotropic salt and added to the glass fiber fleece in the High Pure Spin Filter Tube. Under the buffer conditions used in this process, the plasmid binds to the glass fiber villi, while the contaminants (salts, proteins, and other cellular contaminants) are not. Short washing and spinning steps can easily remove these contaminants. After purification, the plasmid can be easily eluted in a small amount of low-salt buffer or water.

Purity or Quality Grade
Plasmid pUC19 (4.5 μg) was purified from 1.5 mL of E. coli JM83 suspension and grown in LB medium containing ampicillin for 16 hours at a cell density of 5 A 600 units/mL. 1 μg of purified plasmid DNA was incubated with 5 units of restriction enzyme Eco RI at +37°C for 1 hour, and then analyzed by agarose gel electrophoresis. The sensitivity of the isolated plasmid DNA to restriction endonuclease digestion is the same as that of plasmid DNA separated by CsCl density centrifugation.

Sample Type (General)
0.5-4.0 mL E. coli culture (harvest density 1.5-5.0 A 600 units/mL)

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For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.

CAR-Manu™ High Pure Plasmid Isolation Kit (CAR-K-Z021)

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