Team:University College London/Protocols/puppy

From 2012.igem.org

Revision as of 20:26, 26 September 2012 by Bayke5 (Talk | contribs)
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)
Incubate all three restriction digest reactions at 37°C for 10 minutes and then heat inactivate at 80°C for 20 minutes. Ligate the Upstream and Downstream Parts into the digested Destination Plasmid. {| class="bigtable"
Destination Plasmid DNA 500 ng
EcoRI-HF 1 µl
PstI 1 µl
10X NEBuffer 2.5 µl
100X BSA 0.5 µl
H2O to 50 µl
Upstream Part digestion 2 µl
Destination Plasmid digestion 2 µl
10X T4 DNA Ligase Buffer 2 µl
T4 DNA Ligase 2 µl
H2O 11 µl