Team:Paris/August 27
From 2008.igem.org
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- | ===Protocol of ligation=== | + | ===Protocol of ligation L171=== |
* 2 µL Ligase Buffer 10X | * 2 µL Ligase Buffer 10X | ||
- | * | + | * 1.5 µL D 189 (vector) |
- | * | + | * 5 µL D 188 (insert) |
- | * | + | * 11.5 µL pure Water (qsp 20 µL) |
* 1 µL T4 DNA ligase at 400 000 U/mL concentration | * 1 µL T4 DNA ligase at 400 000 U/mL concentration | ||
* O/N at 16°C | * O/N at 16°C |
Revision as of 16:33, 27 August 2008
Construction of pFlhB - mRFP Tripart (LVA+)Aim : Construction of "pFlhB-RBS-mRFP-dbl ter" (pFlhB-I732078) We can only do the construction with mRFP Tripart (LVA+) because the stable strain with the Biobricks I732011 (mRFP Tripart LVA-) don't to growth.
DigestionMeasurement of the concentration of D187 purifiedProtocol (it's same that for Miniprep)
Ligation
Cloning of EnvZ* in pSB1A2Transformation results
PCR screening
Cloning of OmpR*DigestionDetermination of the concentration of DNAWe used the biophotometer
Name of the digestions
Protocol of digestion
Cleaning of the digestion productsLigationDetermination of the concentration of DNAWe used the biophotometer
Protocol of ligation L171
Checking mutagenesis FliAFor this, i digested mutated FliA and non-mutated FliA with EcoRI and PstI and put in migration the digestion products running on gels. |