Description
General description
Click here to view the kit protocol
The GenElute Mammalian Genomic DNA Purification Kit provides a simple and convenient method to isolate pure, high molecular weight DNA from a variety of mammalian sources. These kits use a silica-based membrane, specially selected for genomic DNA purification, in a convenient spin column format.
The starting materials are lysed in a chaotropic salt-containing solution to ensure the thorough denaturation of macromolecules. The addition of ethanol causes DNA to bind when the lysate is spun through a silica membrane in a microcentrifuge tube. After washing to remove the contaminants, the DNA is eluted in 200 μL of a Tris-EDTA solution. The expected yields of genomic DNA will vary depending on the amount and type of starting material used. DNA purified with the kit has an A₂₆₀/A₂₈₀ ratio between 1.6 and 1.9 and can be up to 50 kb in length.
The purified genomic DNA is ready for immediate use in downstream applications such as restriction digest, PCR, southern blots, and sequencing reactions.
Application
The purified genomic DNA is ready for immediate use in downstream applications such as:
restriction endonuclease digestions
PCR
Southern blots
sequencing reactions
cloning
ligation
Biochem/physiol Actions
The starting materials are lysed in a chaotropic salt-containing solution to ensure the thorough denaturation of macromolecules. Addition of ethanol causes DNA to bind when the lysate is spun through a silica membrane in a microcentrifuge tube. After washing to remove the contaminants, the DNA is eluted in 200 μL of a Tris-EDTA solution. The expected yields of genomic DNA will vary depending on the amount and type of starting material used. DNA purified with the kit has an A₂₆₀/A₂₈₀ ratio between 1.6 and 1.9 and can be up to 50 kb in length.
Features and Benefits
Expected yield: 25 μg from 2 x 10⁶ cultured cells; 30 μg from 25 mg of tissue
Elution volume: 200 - 400 μl
Time required: 20 min after lysis
A₂₆₀/A₂₈₀ ratio: 1.6 - 1.9
Mechanical homogenization required: No
Other Notes
For additional information, please see www.sigma-aldrich.com/genomicdna.
Legal Information
GenElute is a trademark of Sigma-Aldrich Co. LLC
