Paris/July 17

From 2007.igem.org

(Difference between revisions)
(→PCRs)
Line 4: Line 4:
* The Lox66-DapAColi PCRs worked very well in all conditions
* The Lox66-DapAColi PCRs worked very well in all conditions
=> Gel purification
=> Gel purification
 +
 +
{{Paris_PCR| Title = FtsZ-E.Coli
 +
|Name= Ftsz-E.Coli
 +
|Annealing= 55°C
 +
|Elongation= 2m30'
 +
|Cycles= 30
 +
|Buffer=  5x 10µL
 +
|MgCl2= 10µM 0µL
 +
|dNTP= 10µM 1µL
 +
|n_oligoF= 1 Lox71-FtsZ-F
 +
|v_oligoF= 10µM 2.5µL
 +
|n_oligoR= 2 FtsZ-R
 +
|v_oligoR= 10µM 2.5µL
 +
|water= 34µL
 +
|pol= Phusion 0.5µL
 +
|DNA= toothpick in glycerol stock of MG1655
 +
|}}
 +
 +
{{Paris_PCR| Title = DapA-Subtilis
 +
|Name= DapA-Subtilis
 +
|Annealing= 55°C
 +
|Elongation= 2m30'
 +
|Cycles= 30
 +
|Buffer=  5x 10µL
 +
|MgCl2= 10µM 0µL
 +
|dNTP= 10µM 1µL
 +
|n_oligoF= 8 Lox66-DapASubtilis-F
 +
|v_oligoF= 10µM 2.5µL
 +
|n_oligoR= 9 DapASubtilis R
 +
|v_oligoR= 10µM 2.5µL
 +
|water= 34µL
 +
|pol= Phusion 0.5µL
 +
|DNA= 0.5µL of Subtilis DNA from stock (DB tube in fridge)
 +
|}}
 +
 +
{{Paris_PCR| Title = Assembling PCR DGAT
 +
|Name= assembly PCR of DGAT
 +
|Annealing= 55°C
 +
|Elongation= 2m30'
 +
|Cycles= 30
 +
|Buffer=  5x 10µL
 +
|MgCl2= 10µM 0µL
 +
|dNTP= 10µM 1µL
 +
|n_oligoF=
 +
|v_oligoF= 10µM 2.5µL
 +
|n_oligoR=
 +
|v_oligoR= 10µM 2.5µL
 +
|water= 34µL
 +
|pol= Phusion 0.5µL
 +
|DNA= 1µL of PCR product DGAT 1 1µL of PCR product DGAT 2
 +
|}}

Revision as of 10:34, 17 July 2007

PCRs

  • The assembly PCRs did not work... we'll try them again !
  • The Lox66-DapAColi PCRs worked very well in all conditions

=> Gel purification

PCR : FtsZ-E.Coli
Name Ftsz-E.Coli
Annealing T° 55°C
Time of elongation 2m30'
Number of Cycles 30
Buffer 5x 10µL
MgCl2 10µM 0µL
dNTP 10µM 1µL
oligoF 1 Lox71-FtsZ-F 10µM 2.5µL
oligoR 2 FtsZ-R 10µM 2.5µL
water 34µL
polymerase Phusion 0.5µL
DNA toothpick in glycerol stock of MG1655
PCR : DapA-Subtilis
Name DapA-Subtilis
Annealing T° 55°C
Time of elongation 2m30'
Number of Cycles 30
Buffer 5x 10µL
MgCl2 10µM 0µL
dNTP 10µM 1µL
oligoF 8 Lox66-DapASubtilis-F 10µM 2.5µL
oligoR 9 DapASubtilis R 10µM 2.5µL
water 34µL
polymerase Phusion 0.5µL
DNA 0.5µL of Subtilis DNA from stock (DB tube in fridge)
PCR : Assembling PCR DGAT
Name assembly PCR of DGAT
Annealing T° 55°C
Time of elongation 2m30'
Number of Cycles 30
Buffer 5x 10µL
MgCl2 10µM 0µL
dNTP 10µM 1µL
oligoF 10µM 2.5µL
oligoR 10µM 2.5µL
water 34µL
polymerase Phusion 0.5µL
DNA 1µL of PCR product DGAT 1 1µL of PCR product DGAT 2
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