usage
sufficient for 10 purifications
Quality Level
technique(s)
DNA purification: suitable
test parameters
: 70—120 min kit run time, sample volume: 1.5 mL bacterial culture
storage temp.
15-25°C
General description
The GenElute Bacterial Genomic DNA kit provides a simple and convenient way to isolate pure genomic DNA from gram-negative bacteria. For most gram-positive bacteria, the kit must be used in conjunction with the optional lysozyme (L4919) to effectively lyse the thick peptidoglycan cell walls. A Gram-Positive Lysis Solution is provided as a diluent for preparing the lysozyme stock solutions.
The kit combines the advantages of a silica-based system with a microspin format and eliminates the need for expensive resins, alcohol precipitation, and hazardous organic compounds such as phenol and chloroform.
The kit combines the advantages of a silica-based system with a microspin format and eliminates the need for expensive resins, alcohol precipitation, and hazardous organic compounds such as phenol and chloroform.
Application
The purified bacterial genomic DNA is ready for downstream applications such as:
- restriction endonuclease digestions
- PCR
- Southern blots
- cloning
Biochem/physiol Actions
The bacteria are lysed in a chaotropic salt-containing solution to ensure the thorough denaturation of macromolecules. The addition of ethanol causes the DNA to bind when the lysate is spun through a silica membrane into a microcentrifuge tube. After washing to remove the contaminants, the DNA is eluted in 200 μL of a Tris-EDTA solution.
The expected yield of genomic DNA will vary depending on the cell density of the bacterial culture and the bacterial species and strain used. DNA purified with the GenElute kit has an A260/A280 ratio between 1.6 and 1.9 and can be up to 50 kb in length.
The expected yield of genomic DNA will vary depending on the cell density of the bacterial culture and the bacterial species and strain used. DNA purified with the GenElute kit has an A260/A280 ratio between 1.6 and 1.9 and can be up to 50 kb in length.
Features and Benefits
- Starting material: Up to 1.5 mL of culture
- Expected yield: Up to 20 μg
- Elution volume: 400 μl
- Time required: 70 - 120 min
- A260/A280 ratio: 1.6 - 1.9
- No phenol, chloroform, or ethanol precipitation required
- Typical DNA yields of 15 μg - 20 μg
- Protocols provided for Gram + and Gram - bacteria
- High quality genomic DNA in less than 2 hours
- Purified DNA has an A260/A280 ratio between 1.6 and 1.9
Other Notes
For additional information, please see www.sigma-aldrich.com/genomicdna.
Legal Information
GenElute is a trademark of Sigma-Aldrich Co. LLC
signalword
Danger
Hazard Classifications
Acute Tox. 4 Oral - Eye Irrit. 2 - Flam. Liq. 3 - Met. Corr. 1 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
target_organs
Central nervous system, Respiratory system
저장 등급
3 - Flammable liquids
wgk
WGK 3
프로토콜
GenElute™ Bacterial Genomic DNA Kit protocol describes a simple and convenient way for the isolation of pure genomic DNA from bacteria.
Ivan M Mukisa et al.
International journal of food microbiology, 160(1), 1-10 (2012-11-13)
Obushera includes four fermented cereal beverages from Uganda namely: Obutoko, Enturire, Ekitiribita and Obuteire, whose microbial diversity has not hitherto been fully investigated. Knowledge of the microbial diversity and dynamics in these products is crucial for understanding their safety and
Lizbeth Sayavedra et al.
Environmental microbiology, 23(6), 3164-3181 (2021-04-21)
Sulfate-reducing bacteria (SRB) are widespread in human guts, yet their expansion has been linked to colonic diseases. We report the isolation, sequencing and physiological characterization of strain QI0027T , a novel SRB species belonging to the class Desulfovibrionia. Metagenomic sequencing
Pierre Stallforth et al.
Proceedings of the National Academy of Sciences of the United States of America, 110(36), 14528-14533 (2013-07-31)
Stable multipartite mutualistic associations require that all partners benefit. We show that a single mutational step is sufficient to turn a symbiotic bacterium from an inedible but host-beneficial secondary metabolite producer into a host food source. The bacteria's host is