From 2007.igem.org

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	//document.write("insert is v1, and vector is v2");		//document.write("insert is v1, and vector is v2");
-	alert("insert size is " + SI + "(bp)" + " \nand vector size is " + SV + "(bp)");	+	alert(insert size is SI (bp) \n vector size is SV (bp));
	}		}

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Revision as of 07:05, 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)

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

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