Template:Team:KULeuven/29 August 2009/VanillinProduction

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* Split the restriction digest in 2 parts
* Split the restriction digest in 2 parts
-
** 1 Part was directly used for ligation using double the mass required
+
** 1 Part was directly used for ligation using double the mass required(SAMS II and EF II)
{| border ="1" align="center"
{| border ="1" align="center"
!| Part || DNA ng || volume µl
!| Part || DNA ng || volume µl
|- align="center"
|- align="center"
-
| Sam5||100 || 0,8
+
| ''sam5''||100 || 0,8
|- align="center"
|- align="center"
-
| Sam8||126,4 || 2,7
+
| ''sam8''||126,4 || 2,7
|- align="center"
|- align="center"
-
| Ech ||100 || 1
+
| ''ech'' ||100 || 1
|- align="center"
|- align="center"
-
| Fcs ||244 || 2,2
+
| ''fcs'' ||244 || 2,2
|-
|-
|}
|}
-
* The 2nd part was first put on gel electrophorese and purified before it was used for ligation
+
* The 2nd part was first put on a gel electrophoresis and purified before it was used for ligation
 +
* Gel electrophoresis of restriction on ''sam5, sam8, fcs and ech''
 +
** ''sam5'' and ''sam8'' showed expected results
 +
** ''fcs'' and ''ech'' had unexpected signals
 +
***''fcs'' showed only 1 result on gel, expected two
 +
***''ech'' showed 4 results in gel (all too short), expected one
 +
** ''sam5'' and ''sam8'' were purified and ligated (SAMS I)
 +
 
 +
{| border ="1" align="center"
 +
!| Part || DNA conc ng/µl|| 260/280
 +
|- align="center"
 +
| ''sam5''||6,6 || 1,98
 +
|- align="center"
 +
| ''sam8''||4,1 || 2,88
 +
|-
 +
|}
 +
 
 +
* ''ech'' and ''fcs'' were cut again and put overnight (over the weekend...) in 16°C

Latest revision as of 11:43, 14 September 2009

  • Split the restriction digest in 2 parts
    • 1 Part was directly used for ligation using double the mass required(SAMS II and EF II)
Part DNA ng volume µl
sam5100 0,8
sam8126,4 2,7
ech 100 1
fcs 244 2,2
  • The 2nd part was first put on a gel electrophoresis and purified before it was used for ligation
  • Gel electrophoresis of restriction on sam5, sam8, fcs and ech
    • sam5 and sam8 showed expected results
    • fcs and ech had unexpected signals
      • fcs showed only 1 result on gel, expected two
      • ech showed 4 results in gel (all too short), expected one
    • sam5 and sam8 were purified and ligated (SAMS I)
Part DNA conc ng/µl 260/280
sam56,6 1,98
sam84,1 2,88
  • ech and fcs were cut again and put overnight (over the weekend...) in 16°C
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