NYMU Taipei/phptest

From 2007.igem.org

(Difference between revisions)
 
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* formula
* formula
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** '''300''' * SI * 3 * 10 (損失因子) / SV = CI * VI * L /50
+
** '''300''' * SI * 3 (ligation ratio) * 10 (損失因子) / SV = CI * VI * L /50
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** vector weight is '''300''' ng
+
** mass of vector (MV) is '''300''' ng
** SI (bp) is size of insert
** SI (bp) is size of insert
** SV (bp) is size of vector
** SV (bp) is size of vector

Latest revision as of 10:07, 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)
ligation ratio


  • 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)
    • for vector
      • we assume 300 ng for digestion and 90% loss
      • thus, amplify 10X from 300 ng to 3,000 ng = 3 mg
      • 3,000 ng = CV (ug/uL) * 1000 (ng/ug) * DVV, DVV (digested vector volume) = 3/CV

  • total gel separation volume is 50 uL

  • total ligation volume is 10uL
    • (MI/MV) = R * (SI/SV)
    • assume mass of vector (MV) is 300 ng
      • MI = R * (SI/SV) * MV = 300R * (SI/SV) (ng)
    • 10X buffer (1uL)
    • ligase (1uL)
    • VV + L + water (8uL)

  • formula
    • 300 * SI * 3 (ligation ratio) * 10 (損失因子) / SV = CI * VI * L /50
    • mass of vector (MV) 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
    • R is ranged from 1 to 10