Silver: Ligation

Using NEB T4 DNA ligase:
 * Mix:
 * 1) *50-100 ng de-phosphorylated, digested vector DNA
 * 2) *3-10x molar excess digested insert DNA
 * 3) *2 μL 10x T4 DNA ligase buffer
 * 4) *x μL distilled water (final volume of reaction should be 20 μL)
 * 5) *1 μL T4 DNA ligase
 * 6) Mix thoroughly, incubate for 30 minutes at room temperature
 * 7) Proceed to transformation. Store excess ligation mixture at -20 °C

Using Roche's Rapid DNA Ligation Kit, stored at -20 °C. Keep the total mass of DNA below 200 ng.
 * Mix:
 * 1) *50-100 ng de-phosphorylated, digested BioBrick vector (~2 µL, ~18 pmol vector).
 * 2) *4-10x molar excess digested insert DNA.
 * 3) *2 µL tube #2 (DNA dilution buffer)
 * 4) *distilled water to final volume of 10 µL total volume
 * 5) Add 10 µL of freshly mixed tube #1 (2x T4 DNA ligase buffer) or 2uL of 10x T4 DNA ligase buffer
 * 6) Add 1 µL tube #3 (T4 DNA ligase).
 * 7) Mix well, incubate for 15 min. at room temperature.
 * 8) Proceed to transformation. Store excess ligation mixture at -20 °C.