NYMU Taipei/phptest
From 2007.igem.org
(Difference between revisions)
| (One intermediate revision not shown) | |||
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<script language="javascript"> | <script language="javascript"> | ||
| - | function CheckId(SI,SV,CI,CV,VI,L){ | + | function CheckId(SI,SV,CI,CV,VI,L,R){ |
var SI = SI; | var SI = SI; | ||
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var VI = VI; | var VI = VI; | ||
var L = L; | var L = L; | ||
| + | var R = R; | ||
var DVV = 3/CV; | var DVV = 3/CV; | ||
| + | var MI = R * (SI/SV); | ||
//document.write("insert is v1, and vector is v2"); | //document.write("insert is v1, and vector is v2"); | ||
| - | alert("insert size = " + SI + "bp\n" + "vector size = " + SV + "bp\n" + "vector concentration = " + CV + "ug/uL\n" + "digested vector volume = " + DVV + "uL"); | + | alert("insert size = " + SI + "bp\n" + "vector size = " + SV + "bp\n" + "vector concentration = " + CV + "ug/uL\n" + "digested vector volume = " + DVV + "uL\n" + "insert mass = " + MI + "ug"); |
} | } | ||
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<INPUT type="text" name="L"> volume (uL) of insert for ligation (L)<BR> | <INPUT type="text" name="L"> volume (uL) of insert for ligation (L)<BR> | ||
<INPUT type="text" name="R"> ligation ratio<BR> | <INPUT type="text" name="R"> ligation ratio<BR> | ||
| - | <input type=button value="送出" Onclick="CheckId(SI.value, SV.value, CI.value, CV.value, VI.value, L.value);"> | + | <input type=button value="送出" Onclick="CheckId(SI.value, SV.value, CI.value, CV.value, VI.value, L.value, R.value);"> |
</form> | </form> | ||
</body> | </body> | ||
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---- | ---- | ||
* formula | * formula | ||
| - | ** '''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 |
| - | ** vector | + | ** 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 | ||
| Line 63: | Line 65: | ||
** CI (ng/uL) is concentration of insert | ** CI (ng/uL) is concentration of insert | ||
** CV (ng/uL) is concentration of vector | ** CV (ng/uL) is concentration of vector | ||
| + | ** R is ranged from 1 to 10 | ||
Latest revision as of 10:07, 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
- (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
