From 2007.igem.org

(bp) size of insert part (SI)
(bp) size of vector part (SV)
(ug/uL) concentration of insert (CI)
(ug/uL) concentration of vector (CV)
volume (uL) of insert for gel separation (VI)
volume (uL) of insert for ligation (L)

---

• total digestion volume is 20 uL (typical)
  • enzyme 1 and enzyme 2 (1uL * 2)
  • 10X buffer (2uL)
  • BSA (2uL)
  • plasmid + ddH2O (14uL)

---

• total gel separation volume is 50 uL
  • vector weight is 300 ng

---

• formula
  • (1) 300 * SI * 3 * 10 / SV = CI * X * L /50
  • (2) 300 ng = CV (ng/uL) * VV (uL), VV = 300/CV
  • VV is volume of vector