SDU-Denmark/17 July 2010

From 2010.igem.org

Progress Report

The Flagella group
At last, we are moving forward! Yesterday we ran the PCR with Pfu at the three most successful temperatures from the Taq gradient PCR. We wanted to make sure that we would get some sort of results. And that we did! The gel showed clear bands at the appropriate lengths, and the NanoDrop showed high concentrations of DNA.

Because there is a stop-codon within the gene, we want to induce a silent mutation. That has been done by using specially designed primers, which results in two strands of the gene that overlap over the mutated area. The overlapping regions should be able to bind to each other in a new PCR, acting as primers for the opposite strand, and the complete mutated gene should the be transcribed. This PCR reaction will be carried out monday. And should it succeed we will have made our first biobrick, ready to be linked to a promoter and RBS and inserted into a plasmid.

Although that sounds simple, our experience has made us sure that we will run into unforeseen obstacles, so this will probably take us a few weeks to complete. But it's easy to have a positive outlook, when we have just run, what we think is the most beautiful PCR and gel of all time!

--Sheila 08:45, 17 July 2010 (UTC)