This product contains highly efficient competent cells prepared from E. coli DH5α. They can be transformed by supercoiled plasmid DNA (pUC19) with a high efficiency (~1 x 109 cfu/μg). This product is appropriate for plasmid amplification, transformation of ligated DNA, and cloning using the TOPcloner™ TA/Blunt kit (Cat.# EZ001, EZ002).
- Strain : DH5α
- Genotype: F’ Φ80lacZ• ΔM15 •ƒ(lacZYA-argF)U169 deoR recA1 endA1 hsdR17(rk-, mk+) phoA supE44 thi-1 gyrA96 relA1
Applications- Amplification and purification of plasmid DNAs- Cloning with ligated DNA mixtures- Cloning with TOPcloner TA/Blunt kits (Cat.# EZ001, EZ002)- Construction of genomic DNA and cDNA librariesQuality control- Transformation efficiency : ~1 x 109 cfu/μg of pUC19
Transformation of competent cells
Take out SOC media and warm up to room temperature in advance.
→ Thaw a tube containing competent cells on ice.
→ Add 1~10 μl of DNA sample to the tube and mix gently.
→ Leave on ice for 30 min.
→ Apply heat-shock at 42℃ for 30 sec and leave on ice for 2 min.
→ Add 400 μl of SOC media (prewarmed to room temperature) to each tube on a clean bench.
→ Incubate for 1 hr at 37℃ in a shaking incubator.
→ Plate 20-200 μl of the sample on plates (prewarmed to room temperature).
→ Incubate overnight at 37℃. Turn the plates upside down during incubation. Examine colonies on the next day.
- Do not use a pipette when thawing competent cells or mixing DNA samples with cells.
- Do not freeze competent cells once thawed.
- Use X-Gal/ IPTG plates for blue/white colony selection, if needed.