NYMU Taipei/phptest
From 2007.igem.org
(Difference between revisions)
| Line 14: | Line 14: | ||
//document.write("insert is v1, and vector is v2"); | //document.write("insert is v1, and vector is v2"); | ||
| - | alert("insert size = " + SI + " | + | alert("insert size = " + SI + "bp\n" + "vector size = " + SV + "bp\n" + "vector concentration = " + CV + "ug/uL"); |
} | } | ||
Revision as of 09:50, 16 September 2007
- 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
- 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
