Team:Paris/July 17
From 2008.igem.org
(Difference between revisions)
(→Screening PCR of the transformations) |
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- | ==Results of the transformations== | + | |
+ | |||
+ | ==Results of the transformations (Biobricks 2007)== | ||
Legende : | Legende : | ||
- | * few , medium, a lot --> description of the number of CFU(Colony forming unit) produce by the transformation | + | * few , medium, a lot --> description of the number of CFU (Colony forming unit) produce by the transformation |
- | * A: Ampicilin, K: | + | * A: Ampicilin, K: Kanamycin |
Line 20: | Line 22: | ||
|align="center"|positive control | |align="center"|positive control | ||
|align="center"|A | |align="center"|A | ||
- | |align="center"|2. | + | |align="center"|2.800 CFU |
|- | |- | ||
|align="center"| - | |align="center"| - | ||
Line 92: | Line 94: | ||
|align="center"|a lot | |align="center"|a lot | ||
|} | |} | ||
+ | |||
==Cloning of the transformations== | ==Cloning of the transformations== | ||
+ | |||
* Test 8 clones by Biobricks | * Test 8 clones by Biobricks | ||
- | * Incubate O/N at 37°C | + | * Incubate O/N at 37°C on proper plates |
- | |||
+ | ==PCR Screening of the transformants== | ||
- | + | Use of the same 8 clones of Biobricks that have been tested for cloning | |
- | |||
- | |||
- | |||
- | |||
- | + | ===Protocol=== | |
- | ===Protocol== | + | |
* 50µl of Mix PCR by tubes | * 50µl of Mix PCR by tubes | ||
* 1 clone | * 1 clone | ||
- | * | + | * Program : 105°C - for 29 cycles : 95°C; 5' - 95°C; 30" - 55°C; 30" - 72°C; 1,5' |
+ | |||
+ | ===Results=== | ||
+ | |||
+ | * '''PCR 1 : J23109 (A)''' | ||
+ | |||
+ | {|border="1" Paris_PCR_1| Title = J23109 (A) | ||
+ | |'''PCRs setting''' | ||
+ | | | ||
+ | |Quick Load | ||
+ | |25µL | ||
+ | | | ||
+ | | | ||
+ | |Expected size | ||
+ | |- | ||
+ | |Annealing | ||
+ | | | ||
+ | |n_oligoF | ||
+ | |18 VF2 | ||
+ | |v_oligoF | ||
+ | |10µM 1µL | ||
+ | |1000pb | ||
+ | |- | ||
+ | |55°C | ||
+ | | | ||
+ | |n_oligoR | ||
+ | |19 VR | ||
+ | |v_oligoR | ||
+ | |10µM 1µL | ||
+ | |Sucess | ||
+ | |- | ||
+ | |Time Elongation | ||
+ | | | ||
+ | |Water | ||
+ | |23µL | ||
+ | | | ||
+ | | | ||
+ | |Yes | ||
+ | |- | ||
+ | |1m30' | ||
+ | | | ||
+ | |DNA | ||
+ | |Toothpick of J23109 | ||
+ | | | ||
+ | | | ||
+ | |Image see bands 1 to 8 | ||
+ | |- | ||
+ | |Number cycles | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |[[Image:080717-1.png|thumb|J23109]] | ||
+ | |- | ||
+ | |29 | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |Ladder band 0 | ||
+ | |} | ||
+ | |||
+ | |||
+ | * '''PCR 2 : E0030 (K)''' | ||
+ | |||
+ | {|border="1" Paris_PCR_2| Title = E0030 (K) | ||
+ | |'''PCRs setting''' | ||
+ | | | ||
+ | |Quick Load | ||
+ | |25µL | ||
+ | | | ||
+ | | | ||
+ | |Expected size | ||
+ | |- | ||
+ | |Annealing | ||
+ | | | ||
+ | |n_oligoF | ||
+ | |18 VF2 | ||
+ | |v_oligoF | ||
+ | |10µM 1µL | ||
+ | |723pb | ||
+ | |- | ||
+ | |55°C | ||
+ | | | ||
+ | |n_oligoR | ||
+ | |19 VR | ||
+ | |v_oligoR | ||
+ | |10µM 1µL | ||
+ | |Sucess | ||
+ | |- | ||
+ | |Time Elongation | ||
+ | | | ||
+ | |Water | ||
+ | |23µL | ||
+ | | | ||
+ | | | ||
+ | |Yes | ||
+ | |- | ||
+ | |1m30' | ||
+ | | | ||
+ | |DNA | ||
+ | |Toothpick of E0030 | ||
+ | | | ||
+ | | | ||
+ | |Image see bands 9 to 16 | ||
+ | |- | ||
+ | |Number cycles | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |[[Image:080717-1.png|thumb|E0030]] | ||
+ | |- | ||
+ | |29 | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |Ladder band 0 | ||
+ | |} | ||
+ | |||
+ | |||
+ | * '''PCR 3 : pSB1A3 (A)''' | ||
+ | |||
+ | {|border="1" Paris_PCR_3| Title = pSB1A3 (A) | ||
+ | |'''PCRs setting''' | ||
+ | | | ||
+ | |Quick Load | ||
+ | |25µL | ||
+ | | | ||
+ | | | ||
+ | |Expected size | ||
+ | |- | ||
+ | |Annealing | ||
+ | | | ||
+ | |n_oligoF | ||
+ | |18 VF2 | ||
+ | |v_oligoF | ||
+ | |10µM 1µL | ||
+ | | | ||
+ | |- | ||
+ | |55°C | ||
+ | | | ||
+ | |n_oligoR | ||
+ | |19 VR | ||
+ | |v_oligoR | ||
+ | |10µM 1µL | ||
+ | |Sucess | ||
+ | |- | ||
+ | |Time Elongation | ||
+ | | | ||
+ | |Water | ||
+ | |23µL | ||
+ | | | ||
+ | | | ||
+ | |Yes | ||
+ | |- | ||
+ | |1m30' | ||
+ | | | ||
+ | |DNA | ||
+ | |Toothpick of pSB1A3 | ||
+ | | | ||
+ | | | ||
+ | |Image see bands 1 to 8 | ||
+ | |- | ||
+ | |Number cycles | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |[[Image:080717-2.png|thumb|pSB1A3]] | ||
+ | |- | ||
+ | |29 | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |Ladder band 0 | ||
+ | |} | ||
+ | |||
+ | |||
+ | * '''PCR 4 : C0179 (A)''' | ||
+ | |||
+ | {|border="1" Paris_PCR_4| Title = C0179 (A) | ||
+ | |'''PCRs setting''' | ||
+ | | | ||
+ | |Quick Load | ||
+ | |25µL | ||
+ | | | ||
+ | | | ||
+ | |Expected size | ||
+ | |- | ||
+ | |Annealing | ||
+ | | | ||
+ | |n_oligoF | ||
+ | |18 VF2 | ||
+ | |v_oligoF | ||
+ | |10µM 1µL | ||
+ | | | ||
+ | |- | ||
+ | |55°C | ||
+ | | | ||
+ | |n_oligoR | ||
+ | |19 VR | ||
+ | |v_oligoR | ||
+ | |10µM 1µL | ||
+ | |Sucess | ||
+ | |- | ||
+ | |Time Elongation | ||
+ | | | ||
+ | |Water | ||
+ | |23µL | ||
+ | | | ||
+ | | | ||
+ | |Yes | ||
+ | |- | ||
+ | |1m30' | ||
+ | | | ||
+ | |DNA | ||
+ | |Toothpick of C0179 | ||
+ | | | ||
+ | | | ||
+ | |Image see bands 9 to 16 | ||
+ | |- | ||
+ | |Number cycles | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |[[Image:080717-2.png|thumb|C0179]] | ||
+ | |- | ||
+ | |29 | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |Ladder band 0 | ||
+ | |} | ||
+ | |||
+ | |||
+ | * '''PCR 5 : C0079 (K)''' | ||
+ | |||
+ | {|border="1" Paris_PCR_5| Title = C0079 (K) | ||
+ | |'''PCRs setting''' | ||
+ | | | ||
+ | |Quick Load | ||
+ | |25µL | ||
+ | | | ||
+ | | | ||
+ | |Expected size | ||
+ | |- | ||
+ | |Annealing | ||
+ | | | ||
+ | |n_oligoF | ||
+ | |18 VF2 | ||
+ | |v_oligoF | ||
+ | |10µM 1µL | ||
+ | |876pb | ||
+ | |- | ||
+ | |55°C | ||
+ | | | ||
+ | |n_oligoR | ||
+ | |19 VR | ||
+ | |v_oligoR | ||
+ | |10µM 1µL | ||
+ | |Sucess | ||
+ | |- | ||
+ | |Time Elongation | ||
+ | | | ||
+ | |Water | ||
+ | |23µL | ||
+ | | | ||
+ | | | ||
+ | |Yes | ||
+ | |- | ||
+ | |1m30' | ||
+ | | | ||
+ | |DNA | ||
+ | |Toothpick of C0079 | ||
+ | | | ||
+ | | | ||
+ | |Image see bands 1 to 8 | ||
+ | |- | ||
+ | |Number cycles | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |[[Image:080717-3.png|thumb|C0079]] | ||
+ | |- | ||
+ | |29 | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |Ladder band 0 | ||
+ | |} | ||
+ | |||
+ | |||
+ | * '''PCR 6 : R0079 (A)''' | ||
+ | |||
+ | {|border="1" Paris_PCR_6| Title = R0079 (A) | ||
+ | |'''PCRs setting''' | ||
+ | | | ||
+ | |Quick Load | ||
+ | |25µL | ||
+ | | | ||
+ | | | ||
+ | |Expected size | ||
+ | |- | ||
+ | |Annealing | ||
+ | | | ||
+ | |n_oligoF | ||
+ | |18 VF2 | ||
+ | |v_oligoF | ||
+ | |10µM 1µL | ||
+ | |277pb | ||
+ | |- | ||
+ | |55°C | ||
+ | | | ||
+ | |n_oligoR | ||
+ | |19 VR | ||
+ | |v_oligoR | ||
+ | |10µM 1µL | ||
+ | |Sucess | ||
+ | |- | ||
+ | |Time Elongation | ||
+ | | | ||
+ | |Water | ||
+ | |23µL | ||
+ | | | ||
+ | | | ||
+ | |Yes | ||
+ | |- | ||
+ | |1m30' | ||
+ | | | ||
+ | |DNA | ||
+ | |Toothpick of R0079 | ||
+ | | | ||
+ | | | ||
+ | |Image see bands 9 to 16 | ||
+ | |- | ||
+ | |Number cycles | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |[[Image:080717-3.png|thumb|R0079]] | ||
+ | |- | ||
+ | |29 | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |Ladder band 0 | ||
+ | |} | ||
+ | |||
+ | |||
+ | |||
+ | * '''PCR 7 : E1010 (K)''' | ||
+ | |||
+ | {|border="1" Paris_PCR_7| Title = E1010 (K) | ||
+ | |'''PCRs setting''' | ||
+ | | | ||
+ | |Quick Load | ||
+ | |25µL | ||
+ | | | ||
+ | | | ||
+ | |Expected size | ||
+ | |- | ||
+ | |Annealing | ||
+ | | | ||
+ | |n_oligoF | ||
+ | |18 VF2 | ||
+ | |v_oligoF | ||
+ | |10µM 1µL | ||
+ | |801pb | ||
+ | |- | ||
+ | |55°C | ||
+ | | | ||
+ | |n_oligoR | ||
+ | |19 VR | ||
+ | |v_oligoR | ||
+ | |10µM 1µL | ||
+ | |Sucess | ||
+ | |- | ||
+ | |Time Elongation | ||
+ | | | ||
+ | |Water | ||
+ | |23µL | ||
+ | | | ||
+ | | | ||
+ | |Yes | ||
+ | |- | ||
+ | |1m30' | ||
+ | | | ||
+ | |DNA | ||
+ | |Toothpick of E1010 | ||
+ | | | ||
+ | | | ||
+ | |Image see bands 1 to 8 | ||
+ | |- | ||
+ | |Number cycles | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |[[Image:080717-4.png|thumb|E1010]] | ||
+ | |- | ||
+ | |29 | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |Ladder band 0 | ||
+ | |} | ||
+ | |||
+ | |||
+ | * '''PCR 8 : B0034 (A)''' | ||
+ | |||
+ | {|border="1" Paris_PCR_8| Title = B0034 (A) | ||
+ | |'''PCRs setting''' | ||
+ | | | ||
+ | |Quick Load | ||
+ | |25µL | ||
+ | | | ||
+ | | | ||
+ | |Expected size | ||
+ | |- | ||
+ | |Annealing | ||
+ | | | ||
+ | |n_oligoF | ||
+ | |18 VF2 | ||
+ | |v_oligoF | ||
+ | |10µM 1µL | ||
+ | | | ||
+ | |- | ||
+ | |55°C | ||
+ | | | ||
+ | |n_oligoR | ||
+ | |19 VR | ||
+ | |v_oligoR | ||
+ | |10µM 1µL | ||
+ | |Sucess | ||
+ | |- | ||
+ | |Time Elongation | ||
+ | | | ||
+ | |Water | ||
+ | |23µL | ||
+ | | | ||
+ | | | ||
+ | |Yes | ||
+ | |- | ||
+ | |1m30' | ||
+ | | | ||
+ | |DNA | ||
+ | |Toothpick of B0034 | ||
+ | | | ||
+ | | | ||
+ | |Image see bands 1 to 7 | ||
+ | |- | ||
+ | |Number cycles | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |[[Image:080717-5.png|thumb|B0034]] | ||
+ | |- | ||
+ | |29 | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |Ladder band 0 | ||
+ | |} | ||
+ | |||
+ | |||
+ | * '''PCR 9 : J23101 (A)''' | ||
+ | |||
+ | {|border="1" Paris_PCR_9| Title = J23101 (A) | ||
+ | |'''PCRs setting''' | ||
+ | | | ||
+ | |Quick Load | ||
+ | |25µL | ||
+ | | | ||
+ | | | ||
+ | |Expected size | ||
+ | |- | ||
+ | |Annealing | ||
+ | | | ||
+ | |n_oligoF | ||
+ | |18 VF2 | ||
+ | |v_oligoF | ||
+ | |10µM 1µL | ||
+ | |1000pb | ||
+ | |- | ||
+ | |55°C | ||
+ | | | ||
+ | |n_oligoR | ||
+ | |19 VR | ||
+ | |v_oligoR | ||
+ | |10µM 1µL | ||
+ | |Sucess | ||
+ | |- | ||
+ | |Time Elongation | ||
+ | | | ||
+ | |Water | ||
+ | |23µL | ||
+ | | | ||
+ | | | ||
+ | |Yes | ||
+ | |- | ||
+ | |1m30' | ||
+ | | | ||
+ | |DNA | ||
+ | |Toothpick of J23101 | ||
+ | | | ||
+ | | | ||
+ | |Image see bands 2 to 7 | ||
+ | |- | ||
+ | |Number cycles | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |[[Image:080717-6.png|thumb|J23101]] | ||
+ | |- | ||
+ | |29 | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |Ladder band 0 | ||
+ | |} | ||
+ | |||
+ | |||
+ | * '''PCR 10 : E0040 (A)''' | ||
+ | |||
+ | {|border="1" Paris_PCR_10| Title = E0040 (A) | ||
+ | |'''PCRs setting''' | ||
+ | | | ||
+ | |Quick Load | ||
+ | |25µL | ||
+ | | | ||
+ | | | ||
+ | |Expected size | ||
+ | |- | ||
+ | |Annealing | ||
+ | | | ||
+ | |n_oligoF | ||
+ | |18 VF2 | ||
+ | |v_oligoF | ||
+ | |10µM 1µL | ||
+ | |840pb | ||
+ | |- | ||
+ | |55°C | ||
+ | | | ||
+ | |n_oligoR | ||
+ | |19 VR | ||
+ | |v_oligoR | ||
+ | |10µM 1µL | ||
+ | |Sucess | ||
+ | |- | ||
+ | |Time Elongation | ||
+ | | | ||
+ | |Water | ||
+ | |23µL | ||
+ | | | ||
+ | | | ||
+ | |Yes | ||
+ | |- | ||
+ | |1m30' | ||
+ | | | ||
+ | |DNA | ||
+ | |Toothpick of E0040 | ||
+ | | | ||
+ | | | ||
+ | |Image see bands 4 to 5 | ||
+ | |- | ||
+ | |Number cycles | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |[[Image:080717-7.png|thumb|E0040]] | ||
+ | |- | ||
+ | |29 | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |Ladder band 0 | ||
+ | |} | ||
+ | |||
+ | |||
+ | |||
+ | * '''PCR 11 : R0040 (A)''' | ||
+ | |||
+ | {|border="1" Paris_PCR_11| Title = R0040 (A) | ||
+ | |'''PCRs setting''' | ||
+ | | | ||
+ | |Quick Load | ||
+ | |25µL | ||
+ | | | ||
+ | | | ||
+ | |Expected size | ||
+ | |- | ||
+ | |Annealing | ||
+ | | | ||
+ | |n_oligoF | ||
+ | |18 VF2 | ||
+ | |v_oligoF | ||
+ | |10µM 1µL | ||
+ | |174pb | ||
+ | |- | ||
+ | |55°C | ||
+ | | | ||
+ | |n_oligoR | ||
+ | |19 VR | ||
+ | |v_oligoR | ||
+ | |10µM 1µL | ||
+ | |Sucess | ||
+ | |- | ||
+ | |Time Elongation | ||
+ | | | ||
+ | |Water | ||
+ | |23µL | ||
+ | | | ||
+ | | | ||
+ | |Yes | ||
+ | |- | ||
+ | |1m30' | ||
+ | | | ||
+ | |DNA | ||
+ | |Toothpick of R0040 | ||
+ | | | ||
+ | | | ||
+ | |Image see bands 7 to 8 | ||
+ | |- | ||
+ | |Number cycles | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |[[Image:080717-7.png|thumb|R0040]] | ||
+ | |- | ||
+ | |29 | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |Ladder band 0 | ||
+ | |} |
Latest revision as of 14:35, 31 July 2008
Results of the transformations (Biobricks 2007)Legende :
Cloning of the transformations
PCR Screening of the transformantsUse of the same 8 clones of Biobricks that have been tested for cloning
Protocol
Results
|