IGEM:University of East Anglia (UEA), Norwich, UK/2009/Notebook/NRP-UEA-Norwich iGEM/2013/07/08
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Today was an important day, where the site-directed mutagenesis reactions were performed on our AntGp Neo plasmid (to fix it by adding an Xba1 restriction site) and J04450 (to add Nde1 restriction site). We used Q5 site-directed mutagenesis kit by New England biolabs, our pre-ordered synthesised oligonucleotides (Pre-Xba1 F/R and J04450-Nde1 F/R primers, and out template DNA to perform the PCR reactions following the kit's protocol. (More details in written lab book).
In addition to the two samples, we ran a negative control in which no DNA was added to Pre-Xba1 F/R primers.
After the PCR reactions were done samples were stored in the freezer for further experiments to confirm mutagenesis was a success.
The poster for the UK meet up was sent for printing today.