Team:Paris/August 27
From 2008.igem.org
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==Ligation== | ==Ligation== | ||
===Determination of the concentration of DNA=== | ===Determination of the concentration of DNA=== | ||
+ | We used the biophotometer | ||
+ | *10 µL of template DNA or 10 µL of EB buffer for th blank | ||
+ | *50 µL of pure water | ||
+ | {|border="1" style="text-align:center" | ||
+ | |- | ||
+ | |'''Template DNA''' | ||
+ | |'''Concentration of DNA''' | ||
+ | |- | ||
+ | |D 188 | ||
+ | |9 µg/mL | ||
+ | |- | ||
+ | |D 189 | ||
+ | |30 µg/mL | ||
+ | |} | ||
+ | |||
===Protocol of ligation=== | ===Protocol of ligation=== | ||
Revision as of 16:32, 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
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. |