NYMU Taipei/phptest

From 2007.igem.org

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<html>
<html>
<body>
<body>
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<php?
 
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echo "test";
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<script language="javascript">
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?>
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function CheckId(SI,SV,CI,CV,VI,L,R){
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test
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var SI = SI;
 +
var SV = SV;
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var CI = CI;
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var CV = CV;
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var VI = VI;
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var L  = L;
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var R  = R;
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var DVV = 3/CV;
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var MI = R * (SI/SV);
 +
 
 +
//document.write("insert is v1, and vector is v2");
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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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}
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</script>
 +
 
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<form name=form1>
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    <INPUT type="text" name="SI"> size (bp) of insert part (SI)<BR>
 +
    <INPUT type="text" name="SV"> size (bp) of vector part (SV)<BR>
 +
    <INPUT type="text" name="CI"> concentration (ug/uL) of insert (CI)<BR>
 +
    <INPUT type="text" name="CV"> concentration (ug/uL) of vector (CV)<BR>
 +
    <INPUT type="text" name="VI"> volume (uL) of insert for gel separation (VI)<BR>
 +
    <INPUT type="text" name="L">  volume (uL) of insert for ligation (L)<BR>
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    <INPUT type="text" name="R">  ligation ratio<BR>
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    <input type=button value="送出" Onclick="CheckId(SI.value, SV.value, CI.value, CV.value, VI.value, L.value, R.value);">
 +
</form>
</body>
</body>
</html>
</html>
 +
----
 +
* total digestion volume is 20 uL (typical)
 +
** enzyme 1 and enzyme 2 (1uL * 2)
 +
** 10X buffer (2uL)
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** BSA (2uL)
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** plasmid + ddH2O (14uL = VI + water, 1 <= VI <= 14)
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** for vector
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*** we assume 300 ng for digestion and 90% loss
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*** thus, amplify 10X from 300 ng to 3,000 ng = 3 mg
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*** 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)
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** assume mass of vector (MV) is 300 ng
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*** MI = R * (SI/SV) * MV = 300R * (SI/SV) (ng)
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** 10X buffer (1uL)
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** ligase (1uL)
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** VV + L + water (8uL)
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----
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* formula
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** '''300''' * SI * 3 (ligation ratio) * 10 (損失因子) / SV = CI * VI * L /50
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** mass of vector (MV) is '''300''' ng
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** SI (bp) is size of insert
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** 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

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
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