From 2007.igem.org
PCRs
| PCR : Lox71-KmR-Lox66
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| PCR Settings
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| Buffer (5x)
| 5x 10µL
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| Expected size
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| Annealing (°C)
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| MgCl2 10µM
| 10µM 0µL
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| 1084
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| 55
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| dNTP 10µM
| 10µM 1µL
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| Success
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| Time Elongation
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| Oligo F 10µM
| 10 Lox71-KmR-F
| 2.5µL
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| ?
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| 3m00'
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| Oligo R 10µM
| 11 Lox66-KmR-R
| 2.5µL
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| Image (click to enlarge)
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| Number cycles
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| Water
| 34µl
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| [[Image:|30px]]
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| 35
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| Polymerase
| Phusion 0.5µL
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| Band (0=ladder)
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| DNA
| MP3_pSB1AK3-BBa_B0015 0.5µl
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| PCR : Lox71-FtsA-FtsZ
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| PCR Settings
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| Buffer (5x)
| 5x 10µL
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| Expected size
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| Annealing (°C)
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| MgCl2 10µM
| 10µM 0µL
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| 2588
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| 55
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| dNTP 10µM
| 10µM 1µL
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| Success
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| Time Elongation
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| Oligo F 10µM
| 3 Lox71-FtsA-F
| 2.5µL
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| ? The PCR seemed to have worked but the purification failed
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| 3m00'
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| Oligo R 10µM
| 2 FtsZ-R
| 2.5µL
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| Image (click to enlarge)
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| Number cycles
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| Water
| 34µl
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| [[Image:|30px]]
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| 35
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| Polymerase
| Phusion 0.5µL
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| Band (0=ladder)
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| DNA
| toothpick in MG1655 glycerol
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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
E.Coli pKS::DGAT
We look under microscopy 36 hours after incubation of E.coli transformed by pKS::DGAT and the control E.coli transformed by part B0015 on different LB medium (See July 18).
We can observe colonies (X25).
