From 2007.igem.org

Revision as of 07:18, 16 September 2007 (view source) Blent (Talk | contribs) ← Older edit		Revision as of 09:43, 16 September 2007 (view source) Blent (Talk | contribs) Newer edit →
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	** BSA (2uL)		** BSA (2uL)
	** plasmid + ddH2O (14uL = VI + water, 1 <= VI <= 14)		** plasmid + ddH2O (14uL = VI + water, 1 <= VI <= 14)
-	** (2) 300 ng = CV (ug/uL) * VV (uL) * 1000, VV = 0.3/CV	+	** 300 ng = CV (ug/uL) * VV (uL) * 1000, VV = 0.3/CV
	----		----
	* total gel separation volume is 50 uL		* total gel separation volume is 50 uL
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	----		----
	* formula		* formula
-	** (1) 300 * SI * 3 * 10 (損失因子) / SV = CI * VI * L /50	+	** '''300''' * SI * 3 * 10 (損失因子) / SV = CI * VI * L /50
-		+	** vector weight is '''300''' ng
		+	** SI (bp) is size of insert
		+	** SV (bp) is size of vector
		+	** VI (uL) is volume of insert
	** VV (uL) is volume of vector		** VV (uL) is volume of vector
		+	** CI (ng/uL) is concentration of insert
	** CV (ng/uL) is concentration of vector		** CV (ng/uL) is concentration of vector

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Revision as of 09:43, 16 September 2007

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)
  • 300 ng = CV (ug/uL) * VV (uL) * 1000, VV = 0.3/CV

---

• total gel separation volume is 50 uL

---

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

---

• formula
  • 300 * SI * 3 * 10 (損失因子) / SV = CI * VI * L /50
  • vector weight is 300 ng
  • SI (bp) is size of insert
  • SV (bp) is size of vector
  • VI (uL) is volume of insert
  • VV (uL) is volume of vector
  • CI (ng/uL) is concentration of insert
  • CV (ng/uL) is concentration of vector