Bio-Rad Laboratories, Inc.
2000 Alfred Nobel Dr.Hercules, CA 94547 USA(510) 741-1000
1-800-424-6723
4110111 Rev C
Page 1: ...Bio Rad Laboratories Inc 2000 Alfred Nobel Dr Hercules CA 94547 USA 510 741 1000 1 800 424 6723 4110111 Rev C...
Page 2: ...Aurum Plasmid Mini Kit Instruction Manual For technical service call your local Bio Rad office or in the US call 1 800 424 6723...
Page 3: ...1 Section 2 Kit Components 1 Section 3 Storage Conditions 2 Section 4 Necessary Supplies 2 Section 5 Guidelines for Using the Aurum Plasmid Mini Kit 3 Section 6 Protocol 5 Section 7 Troubleshooting Gu...
Page 4: ...lasmid mini kit is eluted into a small volume of aqueous buffer and is free of salts bacterial chromosomal DNA and RNA The exceptional purity of the plasmid produced by this system makes it ideal for...
Page 5: ...ecessary Supplies Equipment and supplies to be provided by the customer 1 5 2 0 ml capped microcentrifuge tubes 200 95 100 ethanol or reagent grade denatured ethanol 100 ml Microcentrifuge 12 000 x g...
Page 6: ...rmance LB or LBG is recommended for most strains of E coli TB Terrific Broth cultures generally produce lower plasmid yields of more variable quality and are therefore not recommended For optimum plas...
Page 7: ...uired for plasmid purification use the following equation OD600 of undiluted culture x culture volume in ml OD ml For example 12 OD ml of bacteria would require 2 ml of an undiluted culture with an OD...
Page 8: ...commodate 1 5 ml and 2 0 ml tubes Please read the section Guidelines for Using the Aurum Plasmid Mini Kit before proceeding 1 Transfer up to 12 OD ml of plasmid containing bacterial host to a 1 5 2 0...
Page 9: ...ini kit Fig 3b Connection of Aurum mini column to column adaptor plate Aurum mini column Column adaptor plate Manifold top Waste collection tray A stage Manifold base Fig 4 Vacuum setup conditions Vac...
Page 10: ...il all of the lysate has passed through the column Open the vacuum regulator until the gauge indicates 0 Hg 8 The wash solution is supplied as a 5x concentrate Add 4 volumes 100 ml of 95 100 ethanol o...
Page 11: ...cuum off 10 Add 750 l wash solution and reapply vacuum until all liquid has passed through column 11 Transfer mini spin vac column to a 2 ml wash tube Spin 1 min to remove residual wash Collection of...
Page 12: ...fuge the neutralized lysate for 5 min A compact white debris pellet will form along the side or at the bottom of the tube The supernatant or cleared lysate contains the plasmid DNA 6 While centrifugin...
Page 13: ...t flow through 10 Add 750 l wash solution and centrifuge 1 min Decant flow through 11 Centrifuge additional 1 min to remove residual wash solution Collection of Purified Samples 12 Transfer mini spin...
Page 14: ...mes for culture depending upon host broth etc Check age and concentration of antibiotic Excessive amount of Determine OD600 of bacteria processed culture and reduce OD ml of bacteria processed to ensu...
Page 15: ...RNase Replace kit activity due to age or storage conditions Low A260 280 Excessive amount of Determine OD600 of bacteria processed culture and do not exceed 12 OD ml of bacteria processed Incomplete s...
Page 16: ...acterial Use endA host such as degraded host JM109 DH5a or XL1 Blue Plasmid prep Plasmid nicked degraded Check integrity of plasmid performs poorly in or denatured on an analytical gel enzymatic react...
