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 Highly efficient competent cells and protein extraction buffer. Highly Efficient Competent CellsCatalog #: BPS-20000 Size: 20 x 50 µL Description: Quickgrow™ E.coli is specifically selected for its fast growth property. Unlike some other commercial E. coli strains, which take at least 14 hrs for the colonies big enough for picking, Quickgrow™ E.coli can grow to pickable colonies on LB plates within 9 hours. Quickgrow™ E.coli provides α-complementation of the β-galactosidase gene for color selection, may be used for producing single-stranded DNA from M13 or phagemid and may also be used for inducible protein expression of those genes under LacUV5 promoter control. Its genotype is F¢ proA+B+ lacIq D(lacZ)M15/ D(lac-proAB) glnV thi-1 D(hsdS-mcrB)5. Components: 20 x 50 µl cells, 10 ul pUC18 (0.1ng/µl), 10 ml SOC Efficiency: 1.0 x 109 CPU (colony per ug pUC18) Shelf life: 6 months from receipt date Test conditions: Transformations are performed with 50 aliquots of cells and 2 ul of pUC18 control plasmid (10 pg/ul) following the provided protocol. Dilute the reaction 1:100 with RT SOC and 100 ul of the dilution is plated in duplicate on LB agar plates with 50 mg/ml ampicillin. The plates are incubated at 37°C overnight and the efficiency is calculated based on the average number of colonies per plate. Shipping conditions: On dry ice. Storage conditions: –80°C freezer. Click here to download product PDF and Protocol Bacterial Protein Extraction BufferCatalog #: BPS-20010 and BPS-20020 Description and Application: BPS Bacterial Protein extraction Buffer enables extraction of both soluble and insoluble (inclusion bodies) proteins from bacteria. The buffer provides high efficiency recovery of over-expressed proteins and is fully compatible with HIS/GST-tag purification procedures and standard protein concentration determination assays. The buffer can also be used for protein extraction from insect cells. Fomulated in: 20 mM Tris-HCl, pH 7.0 with a non-ionic detergent Stability: >12 months at room temperature Procedure (example for 250mL bacterial culture) 1. Pellet bacterial cells from a 250 mL culture at 5,000 g for 10 minutes. Note: Extraction is typically more effective by freezing/thawing the cell pellet prior proceeding to the next step. 2. Resuspend the pellet in 10 ml of extraction buffer and pipette up and down until the suspension is homogenous. Note: If desired, protease inhibitors can be added to the suspension. 3. Freshly prepare a 20 mg/ml stock solution of lysosyme in BPS Bacterial Protein extraction Buffer. Add 200 μl of the lysozyme stock solution to the cell suspension for a final concentration of 0.4 mg/ml. Incubate on ice for 30 min with gentle shaking of the suspension every 10 minutes. Note: If sample becomes viscous, use DNaseI or sonicate the cell suspension (sonication is recommended) until the suspension becomes fluid enough to ensure an efficient centrifugation. 4. Centrifuge the suspension at 12,500g for 30 minutes and collect the supernatant. Product Listing
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