Paris/July 31

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< Paris(Difference between revisions)
(→PCR mutagenesis DGAT)
 
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[[Paris/July 30|yesterday]] -- [[Paris/August 1|tomorrow]] <br>
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== pTOPO cloning ==
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Cloning of PCR  products: P5, P6, P7, P8, P9, P10 (see [[Paris/Freezer]])
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== Transformation ==
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Ligations from yesterday
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== PCR mutagenesis DGAT ==
== PCR mutagenesis DGAT ==
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|Success= YES
|Success= YES
|Image= DGAT107312007.jpg
|Image= DGAT107312007.jpg
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|Band= 4, 5, 6, 7
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|Band= 3, 4, 5, 6 (11 = control without DNA matrix)
|}}
|}}
<br>
<br>
Line 41: Line 52:
|Success= YES
|Success= YES
|Image= DGAT107312007.jpg
|Image= DGAT107312007.jpg
-
|Band= 8,9,10,11
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|Band= 7,8,9,10 (11 = control without DNA matrix)
|}}
|}}
<br>
<br>

Latest revision as of 17:52, 7 October 2007

yesterday -- tomorrow

pTOPO cloning

Cloning of PCR products: P5, P6, P7, P8, P9, P10 (see Paris/Freezer)


Transformation

Ligations from yesterday


PCR mutagenesis DGAT

DGAT has Pst1 restriction site. We want to mutagenize it to do a biobrick.

PCR : DGAT1
PCR Settings Buffer (5x) 5x 10µL Expected size
Annealing (°C) MgCl2 10µM 10µM 0µL 1087
50 dNTP 10µM 10µM 1µL Success
Time Elongation Oligo F 10µM 27 ForDGAT1 2.5µL YES
2m00' Oligo R 10µM 13 DPst1-DGAT-R 2.5µL Image (click to enlarge)
Number cycles Water 33µl DGAT107312007.jpg
35 Polymerase Phusion 0.5µL Band (0=ladder)
DNA Miniprep pKS::DGAT (1µL = 138ng) 3, 4, 5, 6 (11 = control without DNA matrix)


PCR : DGAT2
PCR Settings Buffer (5x) 5x 10µL Expected size
Annealing (°C) MgCl2 10µM 10µM 0µL 396
50 dNTP 10µM 10µM 1µL Success
Time Elongation Oligo F 10µM 12 DPst1-DGAT 2.5µL YES
2m00' Oligo R 10µM 27 Rev-DGAT1 2.5µL Image (click to enlarge)
Number cycles Water 33µl DGAT107312007.jpg
35 Polymerase Phusion 0.5µL Band (0=ladder)
DNA Miniprep pKS::DGAT (1µL = 138ng) 7,8,9,10 (11 = control without DNA matrix)


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