Solution reagent kit E.Z.N.A.®

for PCRfor nucleic acidsbacteria

Isolate low endotoxin (<1EU/µg) plasmid DNA from up to 5 mL bacterial cultures using spin mini columns Overview Plasmid isolated with traditional purification procedures normally contain high levels of endotoxins (also known as lipopolysaccharides or LPS) that can significantly interfere with transfection experiments downstream. Up to 5 mL of overnight cultures can be used for high copy number plasmids and 10 mL for low copy number. The E.Z.N.A.® Endo-Free Plasmid Mini Kit I integrates an efficient endotoxin removal step into the plasmid purification procedure to produce high-quality transfection grade (<0.1 EU/µg) plasmid for efficient transfection. The bacterial cells are lysed using the alkaline-SDS lysis method. The cleared cell lysate is then treated with ETR reagent to efficiently remove the endotoxins. After adjusting the binding conditions, the cell lysate is applied into the HiBind® DNA column and purified DNA is eluted from the column membrane. Rapid – 30 minutes or less Safe – No phenol/chloroform extractions Versatile – Spin and vacuum formats available High-quality – Endotoxins efficiently reduced (<0.1 EU/µg) Specifications FEATURES - SPECIFICATIONS Downstream Application - Sensitive cell line transfection; gene therapy etc. Starting Material - 1-5 mL LB culture with OD600 between 2 and 3; or equivalent Plasmid Type - High-copy, low-copy, cosmid DNA Processing mode - Manual, centrifugation Throughput - 1 - 24 DNA binding technology - Silica mini spin column Lysate clearing method - Centrifugation Processing Time - <30 minutes Yield - 15-25 µg for high copy-number; 0.1-5 µg for low copy-number Other - Includes endotoxin removal step