Revision as of 13:45, 24 August 2012 by Gcn10tdu (Talk | contribs)




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 7

Day 1


. Research into sequencing companies to have our BioBricks sequenced.

. Finalising ordering of genes to be synthesised.


. Transformation of CFP, RFP and GFP by Rebecca.

. Plan of requirements for sequencing of B-M and M-B

Day 2


. Message from GenScript confirming synthesising of ordered genes.

' Plates were found to not be contaminated but there was no growth on any of the plates.

. Retransformed CFP, RFP and GFP into alpha competent cells and plated. Positive controls were made. Some of the transformation culture was retained and more LB medium was added and grown overnight.

. Planning of growth study. Growth Study protocol is on the Lab Protocol page

Day 3


. The transformation was successful and therefore, inoculations of CFP, RFP and GFP were made.

. Redid the growth study using Alpha cells, PyeaR transformed cells, B-M transformed cells and M-B transformed cells. The experiment lasted 12 hours to give a representative view.

. Restriction digest of B-M and M-B using Pst1 and Spe1.

. Minipreps of M-B and B-M.

Day 4


. Minipreps of CFP, RFP and GFP.

. Ran the restriction digest on a gel.

. Ran a lag phase only growth study which involved starting the cultures at a lower absorbance and hence concentration.

. Arrival of primers.

. iGEM PCR primer stock made up to 100µM, then further diluted.

. Preparation of plasmid DNA of our BioBricks to be sent for sequencing. 6µL of the DNA was placed into separate eppendorfs to be sent off.

Day 5


Rachel carried out a gel purification (on both MB and BM from the gel run) using promega wizzard kit (Ref: A9281), and following their protocal.