Template:Team:KULeuven/14 August 2009/VanillinProduction

From 2009.igem.org

(Difference between revisions)
 
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# Gel extraction of the 4 lanes from Sam8 ({{kulpart|BBa_I742106}}) and ech ({{kulpart|BBa_I742113 }})
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# Gel extraction of the 4 lanes from ''sam8'' ({{kulpart|BBa_I742106}}) and ''ech'' ({{kulpart|BBa_I742113 }})
#* To test an additional blanc, we extracted a blanc piece of gel
#* To test an additional blanc, we extracted a blanc piece of gel
#* Nanodrop results were done with the conventional blanc (just elution buffer) and with the extracted blanco gel. However, since the results between both varied too much and the extracted blanco gel was not significantly better, we decided to use the conventional blanco in order to compare between former nanodrop measurements
#* Nanodrop results were done with the conventional blanc (just elution buffer) and with the extracted blanco gel. However, since the results between both varied too much and the extracted blanco gel was not significantly better, we decided to use the conventional blanco in order to compare between former nanodrop measurements
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!| Part || concentration (ng/μl) || 260/280 λ || 260/230 λ
!| Part || concentration (ng/μl) || 260/280 λ || 260/230 λ
|- align="center"
|- align="center"
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|Sam8 ||9,4 || 2,18 ||  
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|''sam8'' ||9,4 || 2,18 ||  
|- align="center"
|- align="center"
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| Ech || 13,6|| 1,94 ||   
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| ''ech'' || 13,6|| 1,94 ||   
|-
|-
|}
|}
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* Ligation of Sam5 (from wednesday) + Sam8 (extracted today)
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* Ligation of ''sam5'' (from Wednesday) + ''sam8'' (extracted today)
  {| border ="1" align="center"
  {| border ="1" align="center"
!| vector || insert
!| vector || insert
|- align="center"
|- align="center"
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| Sam5 || Sam8
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| ''sam5'' || ''sam8''
|- align="center"
|- align="center"
| 3637 bp || 1550 bp
| 3637 bp || 1550 bp
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|-
|-
|}
|}
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* Ligation of ech and fcs. For fcs we took the digest from last week (C1), because this week's restriction digest had a very low concentration (4,2) and a very high 260/280 (6,08)
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* Ligation of ''ech'' and ''fcs''. For ''fcs'' we took the digest from last week (C1), because this week's restriction digest had a very low concentration (4,2) and a very high 260/280 (6,08)
  {| border ="1" align="center"
  {| border ="1" align="center"
!| vector || insert
!| vector || insert
|- align="center"
|- align="center"
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| ech || fcs
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| ''ech'' || ''fcs''
|- align="center"
|- align="center"
| 2929 bp || 1789 bp
| 2929 bp || 1789 bp
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|-
|-
|}
|}
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* Cells were stored in the 16°C
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* Cells were stored at 16°C
* Plated the electroporated cells from the A and C ligation
* Plated the electroporated cells from the A and C ligation
* Made Agar and poured new plates with Ap
* Made Agar and poured new plates with Ap

Latest revision as of 10:04, 14 September 2009

  1. Gel extraction of the 4 lanes from sam8 () and ech ()
    • To test an additional blanc, we extracted a blanc piece of gel
    • Nanodrop results were done with the conventional blanc (just elution buffer) and with the extracted blanco gel. However, since the results between both varied too much and the extracted blanco gel was not significantly better, we decided to use the conventional blanco in order to compare between former nanodrop measurements

Nanodrop:

Part concentration (ng/μl) 260/280 λ 260/230 λ
sam8 9,4 2,18
ech 13,6 1,94
  • Ligation of sam5 (from Wednesday) + sam8 (extracted today)
vector insert
sam5 sam8
3637 bp 1550 bp
50 ng 85,2 ng
7 μl 9 μl
  • Ligation of ech and fcs. For fcs we took the digest from last week (C1), because this week's restriction digest had a very low concentration (4,2) and a very high 260/280 (6,08)
vector insert
ech fcs
2929 bp 1789 bp
50 ng 122 ng
3 μl 9 μl
  • Cells were stored at 16°C
  • Plated the electroporated cells from the A and C ligation
  • Made Agar and poured new plates with Ap
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