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NA2120 Sigma

GenElute Bacterial Genomic DNA Kits

sufficient for 350 purifications

Synonym: Bacterial Genomic DNA, Gen Elute

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Properties

Related Categories DNA & RNA Purification, Genomic DNA Purification, Molecular Biology, from Bacteria
usage   sufficient for 350 purifications
storage temp.   room temp

Description

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.

Other Notes

For additional information, please see www.sigma-aldrich.com/genomicdna.

Application

The purified bacterial genomic DNA is ready for downstream applications such as:
• restriction endonuclease digestions
• PCR
• Southern blots
• cloning

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

Principle

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.

Legal Information

GenElute is a trademark of Sigma-Aldrich Co. LLC

Price and Availability


Microcon Centrifugal Filters

All labs need water

Kit component also available separately

Description

Product #

Add to Cart

Wash Solution 1 SDS W0263
Column Preparation Solution SDS C2112
Proteinase K from Tritirachium album, lyophilized powder, BioUltra, ≥30 units/mg protein, for molecular biology SDS P2308
RNase A solution SDS R6148

Kit component only

Description

Product #

Add to Cart

GramPositive Lysis Solution SDS    
Lysis Solution T SDS    
Lysis Solution C SDS    
Wash Solution Concentrate SDS    
Elution Solution (10 mM Tris-HCl, 0.5 mM EDTA, pH 9.0) SDS    
Nucleic Acid Binding Column SDS    
Safety & Documentation

Safety Information

Symbol 
Signal word 
Danger
Precautionary statements 
RIDADR 
UN 3316 9
Flash Point(F) 
77 °F
Flash Point(C) 
25 °C
Protocols & Articles

Protocols

GenElute Bacterial Genomic DNA Kit Protocol

Sigma's GenElute™ Bacterial Genomic DNA Kit provides a simple and convenient way to isolate pure DNA from a variety of cultured bacteria. The kit contains all of the reagents needed to isolate and pu...
Keywords: Amplification, Anticoagulants, Cell disruption, Centrifugation, Degradations, Digestions, Evaporation, Molecular biology, Nucleic acid denaturation, Phase transitions, Polymerase chain reaction, Precipitation

GenElute Bacterial Genomic DNA Kit, Experienced User Protocol

For experienced users, an abbreviated protocol is listed below. For data, images, troubleshooting and other information.

Peer-Reviewed Papers
15

References

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Description

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459836 Ethyl alcohol, Pure, 200 proof, anhydrous, ≥99.5%
L4919 Lysozyme from chicken egg white, BioUltra, lyophilized powder, ≥98% (SDS-PAGE), ≥40,000 units/mg protein
L7386 Lysostaphin from Staphylococcus staphylolyticus, lyophilized powder, Protein 50-70 % by biuret, ≥500 units/mg protein
M9901 Mutanolysin from Streptomyces globisporus ATCC 21553, lyophilized powder, ≥4000 units/mg protein (biuret), Chromatographically purified
W4502 Water, Molecular Biology Reagent

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Description

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T6400 Tris-Borate-EDTA buffer, 5× Concentrate
R6148 RNase A solution
NA2100 GenElute Bacterial Genomic DNA Kits, sufficient for 10 purifications
NA2110 GenElute Bacterial Genomic DNA Kits, sufficient for 70 purifications
C2112 Column Preparation Solution
W0263 Wash Solution 1
SRE0005 Proteinase K from Tritirachium album, Reagents designed and manufactured under cGMP controls suitable for use in an IVD application.
P2308 Proteinase K from Tritirachium album, lyophilized powder, BioUltra, ≥30 units/mg protein, for molecular biology
E1510 Ethidium bromide solution, BioReagent, for molecular biology, 10 mg/mL in H2O
A9539 Agarose, BioReagent, for molecular biology, low EEO

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