Steps

1. Make sure that the incubator (30/37C) and heat block (42C) are ON.
   1. Put water in the wells of the 42°C heat block.
2. Make sure required antibiotic plates are present.  Make sure you're using the right antibiotic plates for your plasmid's resistance!
   1. Warm plates to 37°C.  Cold plates reduce transformation efficiency by an order of magnitude.
   2. Also warm 500 µl SOC per transformation.
3. Take the DNA out of --20 freezer, let it thaw.
   1. Vortex DNA to mix, then spin down.  Make sure it is completely thawed out!
4. Make sure that all of the required reagents/DNA etc are present at the site of transformation before you take the cells out of the -80.
5. Thaw the competent cells on ice for 3-4 min.
   1. You want to add your DNA right as the last bit of cells' ice melts.  Even if it's still a little slushy, that's okay.
6. Add 1-2 µl of DNA into the comp cells.  Stir with a pipette tip a few times, then put right back on ice.
   1. If you're transforming the result of a reaction (GG, LR, etc) add 1-2 µl of the reaction.  Don't add more: many of these reactions have additives that screws up transformation.
   2. If you're transforming plasmid DNA (from a miniprep) transform only 1 ul at whatever concentration it is.
7. Incubate the cells on ice for 30-40 min.
8. Heat shock the cells for EXACTLY 30 sec at 42 C water bath.
9. Place back on ice for 2 min.
10. Add 450 ul of SOC (37° to RT) medium to each tube (S.O.C is made by dissolving 0.5 ml of 20% glucose in 25 ml of SOB. Make sure that the SOC is clear and not cloudy/ contaminated.)
11. Shake the tubes at 37 C, 280 rpm for 60 min.
12. Plate 100 µl for an LR 25ul for a golden gate, or streak out whatever sticks to a loop for a supercoiled plasmid.
13. Incubate plates upside down overnight at 37 C or 16-18h at 30C.
    Can leave the cells in the incubator for up to 18 hours but no more.