NYMU Taipei/phptest

From 2007.igem.org

(Difference between revisions)
Line 33: Line 33:
** 10X buffer (2uL)
** 10X buffer (2uL)
** BSA (2uL)
** BSA (2uL)
 +
** plasmid + ddH2O (14uL)
 +
----
* total gel separation volume is 50 uL
* total gel separation volume is 50 uL
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Revision as of 06:57, 16 September 2007

(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)

  • vector weight is 300 ng
  • 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
  • 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
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