1. Place the competent cells on ice (i.e. DH5α competent strain, Vazyme, # C502).
2. Pipet 10 μL of the recombination products to 100 μL of the competent cells, flip the tube several times to mix thoroughly (DO NOT VOTEX!), and then place the tube still on ice for 30 min.
▲The volume of transformation products should not be more than 1/6 of the volume of competent cells.
3. Heat-shock the tube at 42 °C for 70 sec and then immediately chill on ice for 2 - 3 min.
4. Add 900 μL of SOC or LB medium (without antibiotics) to the tube. Then, shake at 37 °C for 1 hour at 200 - 250 rpm.
5. Preheat the LB plate which contains appropriate selection antibiotic at 37 °C.
6. Centrifuge the culture at 5,000 rpm for 5 min, discard 900 μL of supernatant. Then, re-suspend the pellet with 100 μL of remaining medium and plate it on an agar plate which contains appropriate selection antibiotic.
7. Incubate at 37 °C for 12 -16 hours.