From 2007.igem.org

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

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• total digestion volume is 20 uL (typical)
  • enzyme 1 and enzyme 2 (1uL * 2)
  • 10X buffer (2uL)
  • BSA (2uL)
  • plasmid + ddH2O (14uL = VI + water, 1 <= VI <= 14)
  • (2) 300 ng = CV (ug/uL) * VV (uL) * 1000, VV = 0.3/CV

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• total gel separation volume is 50 uL

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• total ligation volume is 10uL
  • vector weight is 300 ng
  • 10X buffer (1uL)
  • ligase (1uL)
  • VV + L + water (8uL)

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• formula
  • (1) 300 * SI * 3 * 10 (損失因子) / SV = CI * VI * L /50

• • VV (uL) is volume of vector
  • CV (ng/uL) is concentration of vector