Bacterial Genomic DNA isolation kits are all inclusive ready to use kits for the purification of high-molecular-weight DNA from multiple species of bacteria.

Principle

Bacterial DNA Kits allow rapid and reliable isolation of high-quality total cellular DNA from a wide variety of bacterial species, with the capability to handle up to 1 x 10⁹ bacterial cells. The system combines the reversible nucleic acid-binding properties of HiBind™ matrix with the speed and versatility of spin column technology to yield approximately 15-30g of high purity DNA. The use of mini-columns facilitate the binding, washing, and elution steps thus enabling multiple samples to be simultaneously processed. For more information on columns, click Flexible Multi-Format Columns.

Procedure

Bacterial cells are grown to log phase and harvested by centrifugation. After harvesting the bacterial cell wall is degraded by lysozyme and protease K digestion. Once lysed, solution binding conditions are adjusted to enable efficient DNA membrane binding and the sample is applied to a HiBind^® DNA spin-column. Two rapid wash steps remove trace salt and protein contaminants and in a final step DNA is eluted in sterile water or low ionic strength buffer.

Applications

The DNA isolated using the Bacterial Genomic DNA Isolation Kit is free of all contaminants such as RNA, protein, and metabolites. As a result it ids of the highest purity, with 260/280 absorbance ratios between 1.7 and 1.9, making it well suited for use in the following applications :

• RFLP analysis
• Analysis of gene targeting
• Restriction Digestion
• DNA fingerprinting studies
• Hybridization Techniques
• PCR amplification

Agarose gel analysis of genomic DNA from bacteria using
the Bacterial DNA Kit. Lane M: 1-kb ladder DNA
Lane 1: DH5a, Lane 2: JM 109