Paris/July 20

From 2007.igem.org

(Difference between revisions)
(PCRs)
(PCRs)
Line 3: Line 3:
{{Paris_PCR_0| Title = Lox71-KmR-Lox66
{{Paris_PCR_0| Title = Lox71-KmR-Lox66
-
|Annealing= 55°(3x)-63°(35x)C
+
|Annealing= 55
-
|Elongation= 2m00'
+
|Elongation= 3m00'
-
|Cycles= 38
+
|Cycles= 35
|Buffer=  5x 10µL
|Buffer=  5x 10µL
|MgCl2= 10µM 0µL
|MgCl2= 10µM 0µL
Line 13: Line 13:
|n_oligoR= 11  Lox66-KmR-R
|n_oligoR= 11  Lox66-KmR-R
|v_oligoR= 2.5µL
|v_oligoR= 2.5µL
-
|water= 30µL
+
|water= 34µl
|pol= Phusion 0.5µL
|pol= Phusion 0.5µL
-
|DNA= MP3_pSB1AK3-BBa_B0015 2µL dil.100
+
|DNA= MP3_pSB1AK3-BBa_B0015 0.5µl
-
|Size=
+
|Size= 1084
 +
|Success=
 +
|Image=
 +
|Band=
 +
|}}
 +
 
 +
{{Paris_PCR_0| Title = Lox71-FtsA-FtsZ
 +
|Annealing= 55
 +
|Elongation= 3m00'
 +
|Cycles= 35
 +
|Buffer=  5x 10µL
 +
|MgCl2= 10µM 0µL
 +
|dNTP= 10µM 1µL
 +
|n_oligoF= 3 Lox71-FtsA-F
 +
|v_oligoF= 2.5µL
 +
|n_oligoR= 2 FtsZ-R
 +
|v_oligoR= 2.5µL
 +
|water= 34µl
 +
|pol= Phusion 0.5µL
 +
|DNA= toothpick in MG1655 glycerol
 +
|Size= 2588
|Success=
|Success=
|Image=
|Image=

Revision as of 12:34, 23 July 2007

PCRs

PCR : Lox71-KmR-Lox66
PCR Settings Buffer (5x) 5x 10µL Expected size
Annealing (°C) MgCl2 10µM 10µM 0µL 1084
55 dNTP 10µM 10µM 1µL Success
Time Elongation Oligo F 10µM 10 Lox71-KmR-F 2.5µL
3m00' Oligo R 10µM 11 Lox66-KmR-R 2.5µL Image (click to enlarge)
Number cycles Water 34µl [[Image:|30px]]
35 Polymerase Phusion 0.5µL Band (0=ladder)
DNA MP3_pSB1AK3-BBa_B0015 0.5µl
PCR : Lox71-FtsA-FtsZ
PCR Settings Buffer (5x) 5x 10µL Expected size
Annealing (°C) MgCl2 10µM 10µM 0µL 2588
55 dNTP 10µM 10µM 1µL Success
Time Elongation Oligo F 10µM 3 Lox71-FtsA-F 2.5µL
3m00' Oligo R 10µM 2 FtsZ-R 2.5µL Image (click to enlarge)
Number cycles Water 34µl [[Image:|30px]]
35 Polymerase Phusion 0.5µL Band (0=ladder)
DNA toothpick in MG1655 glycerol

Glycerol stock

  • BBa_pJ23107 clone 3&4 (S14)
  • BBa_I0500 clone 1&2 (S13)

Minipreps on Biobricks

  • BBa_pJ23107 clone 3&4
  • BBa_I0500 clone 1&2