MP Biomedicals™ Rb/CaCl Transformation Salts

Description

SUPER-COMP is an extremely rich medium that significantly increases the competency of the cells when using a simple RbCl/CaCl means of preparing competent host cells.

• Dilute 1mL of a fresh overnight culture of host cells grown in LB medium into 25mL of 37°C SUPER-COMP media in a 500mL flask
• Incubate cells with vigorous aeration for 1-2 hours until cells reach mid-log phase (approximately 8 x 10⁸ cells/mL, OD600 reading 0.8 for most E. coli host cells)
• Centrifuge culture in a sterile tube at 4°C to pellet cells. Resuspend pellet in 15mL of ice cold 1x RbCl/CaCl Transformation Salts
• After 15 min. on ice, centrifuge to pellet cells and gently resuspend in 2mL of ice cold 1x RbCl/CaCl Transformation Salts; Keep cell suspension on ice

Cells are now competent and are immediately ready for DNA uptake. For maximum transformation efficiency, store for 12-24 hours at 4°C to increase competency 4-6 fold (at 48 hours they will be approximately as efficient as when first made). Competent cells can also be frozen in aliquots at -80°C after adding glycerol to 20%.