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Team:Paris Bettencourt/Notebook/Trojan Horse/Thursday 5th September.html
From 2013.igem.org
Trojan Horse
ASDF5th September
Confirmation of silencing (CmR)
Cloning biobricks : ie clone sRNAs (Cm, Kan, lacZ) in pB1C3
|
Reagent |
Volume |
|
Purified PCR Product |
16 ul |
|
H2O |
0 ul |
|
10x FastDigest Buffer |
2 ul |
|
FastDigest Enzyme 1 |
1 ul |
|
FastDigest Enzyme 2 |
1 ul |
|
Total Volume |
20.0 ul |
Digest for 0.5 - 1 hours at 37 °C with shaking
Ligation sRNA into biobrick vector
masse insert = 10*419/2000*100 = 210 ng
Vector : 5 uL
Insert : 10 uL
Buffer : 2 uL
T4 DNA ligase : 1 uL
H2O : 2uL
Let incubate at 22°C for 30 min
Transformation in NEB turbo chemical competent . Plate on Cm
