Ultra BL21 (DE3) pLysS Competent Cells
Contents
1. containing chloramphenicol Equipment and Materials Not Provided 1 SOC medium for recovery 20g I tryptone 5g l yeast extract 10mM NaCl 10mM KCI 10MM MgClo 10mM Warning This product is intended for research use only It is not to be used for diagnostic purposes in humans or animals MgSO 20mM glucose MgCle MgSO and glucose should be added after autoclaving 2 An orbital shaker A 42 C water bath 4 14ml round bottom culture tubes 1 tube per single use aliquot 5 LB agar plates or liquid medium containing the appropriate antibiotic Chloramphenicol 35yg ml in ethanol should be added to plates or liquid medium for selection of the pLysS plasmid Storage Conditions Ultra BL21 DE3 pLysS Competent Cells should be stored in a 80 C freezer Please note that competent cells are very sensitive to cycles of freezing and thawing and should not be exposed to temperature variations Recommended Protocol for Tubes 1 Immediately after taking the tubes from the 80 C freezer place them in ice and wait approximately 5 minutes until they thaw 2 Pipette the DNA to be transformed to the bottom of the tube and mix by pipetting 50l of air to the bottom of the tube Control transformation Dilute pUC19 supercoiled DNA 1 10 with sterile diH2O then add 1 of the diluted pUC19 supercoiled DNA to one of the tubes Discard diluted pUC19 supercoiled DNA after use Note Do not mix by pipetting up and down since tha2. A EdgeBio Ultra BL21 DE3 pLysS Competent Cells Product Cat No Transformations Ultra BL21 DE3 pLysS Competent Cells Tubes 43066 12 Description Ultra BL21 DE3 pLysS Competent Cells are chemically competent and have been manufactured using proprietary technology making the cells highly efficient for DNA uptake thus ultra competent To utilize the cells at their highest efficiency a recommended transformation protocol is included with each kit Ultra BL21 DE3 pLysS Competent Cells are available in single use tubes that provide a simple and reliable method for high efficiency single use transformation of host cells All cells include a test plasmid for quality control purposes Cells are pre dispensed in 50ul aliquots tubes Full processing time for 1 tube including recovery is about 1 hour and 20 minutes to ensure the highest level of transformation Edge BioSystems guarantees transformation efficiencies that exceed 2x10 cfu ug pUC19 Genotype F ompT hsdSg tg mg gal dem DE3 pLysS Cam Kit Components 43066 Ultra BL21 DE3 pLysS Competent Cells 12 tubes pUC19 Supercoiled DNA 100ng ml 1 tube Quality Control Each lot is tested to assure high transformation efficiency using 10pg pUC19 supercoiled DNA and the recommended protocol Transformation efficiency will exceed 2x1 08 cfu ug pUC19 under these conditions The presence of the pLysS plasmid is confirmed by plating the cells in medium
3. t will lower the transformation efficiency 3 Incubate the tubes in ice for 10 minutes 4 Transfer the tubes to a 42 C water bath incubate for 40 seconds and transfer back to ice 5 Incubate the tubes for 2 minutes in ice 6 Transfer the cells into a 14ml round bottom culture tube filled with 1ml of pre warmed SOC medium and then shake at 300 rpm at 37 C for 1 hour 7 Plate cells on pre warmed LB agar selective plates containing chloramphenicol or inoculate into selective liquid medium containing chloramphenicol For the control transformation with pUC19 supercoiled DNA plate 10ul on LB ampicillin agar plates and expect gt 20 colonies gt 2 x 10 cfu ug pUC 19 Page 1 of 2 v 3 Special Note Ultra BL21 DE3 pLysS Competent Cells are based on the T7 expression system This technology was developed at Brookhaven National Laboratory under contract with the U S Department of Energy Consequently U S patents assigned to Brookhaven Science Associates BSA protect this technology These materials are to be used by noncommercial entities for research purposes only Commercial entities require a license from BSA You may refuse these cells by returning the enclosed materials unused To obtain information about licensing please contact the Office of Intellectual Property and Partnerships Brookhaven National Laboratory Building 475D Upton NY 11973 telephone 631 344 7134 or fax 631 344 3729 Warning This prod
4. uct is intended for research use only It is not to be used for diagnostic purposes in humans or animals Page 2 of 2 v 3
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