1. Take an aliquot of chemically competent bacteria from the --80C freezer and thaw on ice. Bacteria must be kept cold at all times until the heat shock, or they will lose competency. Do not leave bacteria thawing for longer that 15 minutes. The bacteria should be good to go after 7 minutes.
2. Add diluted plasmid DNA stock (1-10 ng, e.g. dilute 1 uL of 1 ug/uL plasmid stock into 1 mL of water, and add 1 uL to bacteria) or 5 uL ligation reaction MAX to 100 uL bacteria. Be sure that the volume of DNA solution added to the bacteria doesn’t exceed 5% of the bacterial volume (5 uL for 100 uL bacteria, 10 for 200 uL, etc). You can add more DNA volume (above the 5%) but note that this may decrease the efficiency of your reaction.
3. Incubate bacteria on ice for 30 minutes, then heat shock by placing in 42C water bath for 30 seconds.
4. Place tube on ice for 2 minutes then and add 9x volumes of SOC (prewarmed at 37C). For example, to 100 uL of bacteria you can add 900 uL of SOC.
5. Shake for 60 minutes at 37C (to allow the bacteria to begin expressing antibiotic resistance; you technically don't need to do this for AmpR but you can if you want), and plate on an LB/antibiotic plate. Plates can be placed at 37C prior to the transformation to allow them to dry slightly.