Product Specification
DH5α：100μl/tube
pUC19 (control vector，10pg/μl)：10μl
storage condition(Guarantee period)： -80℃（6 months）
Genotype
F- φ80 lac ZΔM15 Δ(lacZYA-arg F) U169 endA1 recA1 hsdR17(rk-,mk+) supE44λ- thi -1 gyrA96 relA1 phoA
Product Description
DH5α strain is the most common receptor cell in the laboratory. Missing endonuclease (endA) improves the yield and quality of plasmid DNA. Recombinant enzyme deficiency type (recA) reduces the homologous recombination probability of inserted fragments and ensures the stability of inserted DNA. The existence of the lacZ Δ M15 DH5 alpha screening can be used in the blue and white. The >5 x 108 cfu/ CDG DNA was produced by a special process.
Operation Method
1. DH5 alpha cells from - 80 ℃, inserted rapidly in the ice, wait for 5 minutes for the fungus block to melt, add the target DNA (plasmid or connection product) and gently mix the substances at the EP pipe bottom (avoid using the micropiette to mix the substances), ice stand for 25 minutes.
2. Use 42 ℃ water bath to heat shock for 45 seconds, quickly place it back on the ice and let it stand for 2 minutes, shaking will lower conversion efficiency.
3. In centrifuge tube, add 700 μl of sterile medium (2YT or LB) without antibiotic, after blending, 37 ℃, 200 RPM recovery in 60 minutes.
4. Centrifuge at 5000 RPM for one minute to collect bacteria, and use approximately 100μl of supernatant to gently blow the heavy suspension bacteria block and smear it on the 2YT or LB medium containing the corresponding antibiotics.
5. Put in the planar inverted 37 ℃ culture box for the night. If the blue white spot filtering operation, put 37 ℃ to cultivate the tablet at least 15 hours.
Attention
1. The cells in the receptor state had better melt slowly in the ice and insert the target DNA into the ice within 8 minutes.
2. The mixing of plasmids or connecting products should be handled gently.
3. The conversion of high concentration of plasmids or high-efficiency bonding products can reduce the amount of bacteria eventually used for coating.
