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==Day 5==
==Day 5==
. Posters for the Forum returned from the print, ready to start putting them up.
. Russell and Rachel worked on the DNA purification and isolated the plasmids for RFP, eCFP and AraC promoter and put on ice to run on an agarose gel the next day.

Revision as of 08:54, 3 August 2012




Welcome to the NRP UEA iGEM 2012 Wiki Lab Book

Please choose the relevant link to access our diary of that week!

Week Zero | Week One | Week Two | Week Three | Week Four | Week Five | Week Six | Week Seven | Week Eight | Week Nine | Week Ten | Week Eleven | Lab Protocols | Experiments


Week 4

Day 1

Research and Meetings

. Khadija and Pascoe found problem over the weekend with stop codon; two options are scaffold or assembly standard 23. Leaning towards the other assembly standard as it's less risky than the scaffold, however discussion occuring on both. They developed two main constructs and each took one; one saw overlapping stands, the other saw strands together. Khadija produced a logic table of codons that could bind to one another.

. Amy came to visit in order to look at the studio for our film being recorded on Wednesday.


. Joy and Lukas looked at doing the ligation for our first biobrick using pYeaR/CarG promoter.

. Rebecca transformed the ligated biobricks.

. Russell and Rachel left their colonies to grow a little more due to only a few colonies having grown.

Day 2


. Russell and Rachel looked at nitrite reductase sequences.

. The whole team worked with Khadija and Pascoe on the DNA sequence, four working on Khadija's logic map and three working with Pascoe's idea.


. Russell and Rachel's colonies grew and they inoculated 2 tubes of RFP, 2 tubes of CFP and 4 tubes of arabinose promoter; each using a variety of different colonies to maximise effects. They also used 5ul of ampicillin as the antibiotic in the tubes.

. As the plates from yesterday's transformations showed no growth, Rebecca redid two of the transformations, this time with both negative and positive controls. She also made more chlorophenicol plates and inoculated more hybrid promoter transformed E.coli into media culture. The transformations from yesterday were incubated for longer in hope that colonial growth would appear.

. Lukas and Joy isolated plasmid DNA from "E.coli" containing the hybrid promoter 1. They then double digested the plasmid using EcoR1 and Pst1. Sac1 and Spe1 was also used as a test for the correct plasmid isolated. This was also carried out on hybrid promoter 2 plasmids. Following this, they ran the digested plasmid fragments of promoter 1 on gel.

Day 3


. Again the transformation was unsuccessful.

. Rebecca purified hybrid promoter 2 from gel slice.

. Lukas ran hybrid promoter 2 on gel.

Day 4

. Solved the DNA structure for one of the constructs (with two zip regions that join together when transcribed into mRNA)

. Amy and friends visited to work on the video.

Day 5