Description :

Introduction

The EZgene^TM Bacterial gDNA kit provides a rapid and reliable method for isolating high-quality total cellular DNA from a wide variety of bacterial species. Bacterial cells are grown to log-phase and harvested. The bacterial cell wall is digested by lysozyme and Proteinase K. Following lysis, binding conditions are adjusted and the sample is applied to a DNA column.

Key to this kit is our proprietary DNA binding system that allows the high efficient binding of DNA to our ezBind^TM matrix while proteins and other contaminates are removed under certain optimal conditions. Nucleic acids are easily eluted with sterile water or Elution Buffer. Purified DNA is suitable for PCR, restriction digestion, and hybridization techniques. Each DNA column can bind approximately 100 μg genomic DNA. Up to 1 x 10⁹ bacterial cells can be processed per column. The system combines the reversible nucleic acid-binding properties of our ezBind matrix with the speed and versatility of spin column technology to yield up to approximately 15-30 μg of DNA with an OD_260/280 ratio of 1.7-1.9. The kit will isolate all cellular DNA, including plasmid DNA.

Storage and Stability

 The guaranteed shelf life for this kit is 12 months from the date of purchase.

• Once reconstituted in water, Proteinase K and lysozyme must be stored at -20^oC.
• Store RNase A at 4^oC.
• Other components can be stored at 22^oC-25^oC.
• Under cool ambient conditions, a precipitate may form in the Buffer BL/TL. In case of such an event, heat the bottle at 37^oC to dissolve. Store Buffer BL/TL at room temperature.

Kit Contents

Buffer BL contains chaotropic salts, which may form reactive compounds when combines with bleach. Do not add bleach or acidic solutions directly to the preparation waste.

Before Starting

Please go over the handbook and get familiar with the procedure.

•  Prepare a stock solution of Proteinase K (provided) as follows and aliquot into adequate portions. Store aliquots at -20 ^oC. Bring Proteinase K solution to room temperature before use.

•  Prepare a lysozyme stock solution at 50 mg/mL and aliquot into adequate portions. Store each aliquot at -20 ^oC and thaw before use.

•  Add 100% ethanol to DNA Wash Buffer and store at RT

•  Carry out all of centrifugation step at room temperature. Equilibrate Elution Buffer provided to 65^oC.

Materials to be Provided by User

•  Tabletop microcentrifuge and nuclease-free 1.5 mL tubes
•  Water bath set to 30^oC
•  Shaking water bath set to 55^oC
•  Incubator or waterbath set to 65 ^oC
•  Absolute ethanol (96%-100%)

Operating Protocol