Transform Stratagene's XL-10 Gold Ultracompetent cells
Pre-chill 14mL sterile culture tubes on ice.
Pre-heat 0.9mL of NZY+ broth to 42°C.
Thaw XL-10 Gold Ultracompetent cells on ice then mix gently after completely thawed.
Aliquot cells into pre-chilled sterile culture tubes.
Add beta-mercaptoethanol Swirl tube gently then incubate on ice for 10 minutes swirling gently every 2 minutes.
Add 0.1-50ng of DNA or 2uL of ligation product.
Swirl gently then incubate on ice for 30 minutes.
Heat pulse tube at 42°C for exactly 30 seconds.
The duration of the heat pulse is critical.
Incubate cells on ice for 2 minutes.
Add 0.9mL of pre-heated NZY+ broth to each tube.
Incubate at 37°C for 1 hour with shaking at 225-250 rpm.
Plate no more than 200uL of transformation mixture per LB-agar plate with antibiotic selection.
Incubate plates at 37°C overnight.
