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 .
