Team:University College London/Protocols/Week13/2

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Ligations for irrE, nuclease, laccase & curli

Digest upstream part with EcoRI-HF™ and SpeI

Constitutive promoter-rbs construct 500 ng
EcoRI-HF 1µl
Spel 1 µl
10X NEBuffer 2 5 µl
100X BSA 0.5 µl
H2O to 50 µl

Digest downstream part with Xbal and Pstl

Curli (from PCR product)
Laccase (from PCR product)
irrE (from PCR product)
Nuclease (in puc57)
500 ng
Xbal 1 µl
Pstl 1 µl
10X NEBuffer 2 5 µl
100X BSA 0.5 µl
H2O to 50 µl