NYMU Taipei/phptest

From 2007.igem.org

(Difference between revisions)
Line 10: Line 10:
//document.write("insert is v1, and vector is v2");
//document.write("insert is v1, and vector is v2");
-
alert("insert size = " + SI + "(bp)\nand");
+
alert("insert size = " + SI + "(bp)\n");
}
}
Line 36: Line 36:
* total gel separation volume is 50 uL
* total gel separation volume is 50 uL
** vector weight is 300 ng
** vector weight is 300 ng
 +
----
 +
* total ligation volume is 10uL
 +
** 10X buffer (1uL)
 +
** ligase (1uL)
 +
** VV + IV + water (8uL)
----
----
* formula
* formula

Revision as of 07:10, 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)

  • 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)
  • total gel separation volume is 50 uL
    • vector weight is 300 ng
  • total ligation volume is 10uL
    • 10X buffer (1uL)
    • ligase (1uL)
    • VV + IV + water (8uL)
  • 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
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