Microsoft word - ultra bl21 _de3_ plyss competent cells

Ultra BL21 (DE3) pLysS Competent Cells

Product
Transformations
MgSO4, 20mM glucose (MgCl2, MgSO4 and glucose
Description
4. 14ml round bottom culture tubes (1 tube per single use Ultra BL21 (DE3) pLysS Competent Cells are chemically competent and have been manufactured using proprietary 5. LB-agar plates or liquid medium containing the technology making the cells highly efficient for DNA uptake, appropriate antibiotic. Chloramphenicol (35µg/ml in thus ultra competent. To utilize the cells at their highest ethanol) should be added to plates or liquid medium for efficiency, a recommended transformation protocol is Storage Conditions
Ultra BL21 (DE3) pLysS Competent Cells are available in single use tubes that provide a simple and reliable method Ultra BL21 (DE3) pLysS Competent Cells should be stored in for high-efficiency, single use transformation of host cells. All a -80ºC freezer. Please note that competent cells are very cells include a test plasmid for quality control purposes. sensitive to cycles of freezing and thawing and should not be Cells are pre-dispensed in 50µl aliquots (tubes). Full processing time for 1 tube (including recovery) is about 1 hour and 20 minutes to ensure the highest level of Recommended Protocol for Tubes
transformation. Edge BioSystems guarantees transformation efficiencies that exceed 2x108 cfu/µg pUC19. Immediately after taking the tubes from the -80°C freezer, place them in ice and wait approximately 5 Pipette the DNA to be transformed to the bottom of Kit Components 43066
the tube and mix by pipetting 50µl of air to the bottom of the tube. Control transformation: Dilute pUC19 supercoiled DNA 1:10 with sterile diH2O, then add 1µl of the diluted pUC19 supercoiled DNA to one of the tubes. Discard diluted pUC19 supercoiled DNA after Note: Do not mix by pipetting up and down since that
will lower the transformation efficiency.
Incubate the tubes in ice for 10 minutes. Quality Control
Transfer the tubes to a 42°C water bath, incubate for Each lot is tested to assure high transformation efficiency using 10pg pUC19 supercoiled DNA and the recommended protocol. Transformation efficiency will exceed 2x108 cfu/µg pUC19 under these conditions. The presence of the pLysS plasmid is Incubate the tubes for 2 minutes in ice. confirmed by plating the cells in medium containing 6. Transfer the cells into a 14ml round bottom culture tube filled with 1ml of pre-warmed SOC medium and then Equipment and Materials Not Provided
7. Plate cells on pre-warmed LB-agar selective plates containing chloramphenicol or inoculate into selective liquid medium containing chloramphenicol. For the 1. SOC medium for recovery: 20g/l tryptone, 5g/l yeast control transformation with pUC19 supercoiled DNA, extract, 10mM NaCl, 10mM KCl, 10mM MgCl2, 10mM plate 10µl on LB-ampicillin agar plates and expect >20 colonies (>2 x 108 cfu/µg pUC19). Warning: This product is intended for research use only. It is not to be used for diagnostic purposes in humans or animals.
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 product is intended for research use only. It is not to be used for diagnostic purposes in humans or animals.

Source: http://www.edgebio.com/sites/default/files/43066.pdf