Heat Shock Transformation of Chemically Competent Cells

Reagents and Materials
Competent cells
DNA or Ligation reaction
Sterile tubes
42°C Water Bath
LB Plates with appropriate selective antibiotics
SOC medium:

0.5% yeast extract
2% Tryptone
10 mM NaCl
2.5 mM KCl
10 mM MgCl2
10 mM MgSO4
20 mM Glucose

1. Thaw 100ul of cells per transformation on ice. Aliquot to sterile tubes, if necessary.
2. Add 1-10 ng DNA or 1/5 volume of a DNA Ligation reaction to each tube. Incubate on ice for 30 minutes.
3. Heat Shock tubes in a 42°C water bath for 45 seconds.
4. Return tubes to ice for 2 minutes.
5. Add 900ul SOC to each tube and shake at 200 rpm for 1 hour at 37°C.
6. Plate 1ul, 10ul and 100ul onto LB agar containing selective antibiotics. Incubate at 37ºC overnight (16 hours).