Roche 11754777001 1 kit-HIGH PURE PLASMID ISOLATIONKIT

ApplicationUse this High Pure kit to prepare up to 15 g purified pla

Application
Use this High Pure kit to prepare up to 15 µg purified plasmid DNA from bacterial cultures, so it may be used directly for:
PCR
Sequencing
Cloning
In vitro transcription
Generation of labelled hybridization probes
Background Information
Nucleic acids (NA) bind to the surface of glass fiber fleece in the presence of chaotropic salt.  This allows the High Pure filter tube to specifically immobilize plasmid DNA while it is freed of contaminants.
Contents
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, 2 ml 
Principle
Alkaline lysis releases plasmid DNA from bacteria and RNase removes all RNA in the lysate. After cellular debris and (entrapped) genomic DNA are removed by centrifugation, the remaining supernatant is mixed with a chaotropic salt and applied to the glass fiber fleece in a High Pure Spin Filter Tube.
Under the buffer conditions used in the procedure, the plasmid binds to the glass fleece in the High Pure tube, while contaminating substances (salts, proteins and other cellular contaminants) do not. Brief wash-and-spin steps readily remove these contaminants. Once purified, the plasmid can be easily eluted in a small volume of low salt buffer.
Benefits
Quickly purifies up to 24 plasmid samples in <30 min
Reduces handling steps, minimizing the loss and fragmentation of DNA
Makes DNA suitable for immediate downstream processing
Allows quantitative removal of contaminants, to improve the reliability and reproducibility of downstream procedures
Product Description
Sample: 0.5 - 4.0 ml cultures of E. coli (harvested at a density of 1.5 - 5.0 A600 units per ml)  

 

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