Team:NRP-UEA-Norwich/Week6

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NRP UEA iGEM 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

Contents

Week 6

Day 1

Labs

. We have BioBricks! The plates containing the ligations between B-M and M-B to pSB1C3 showed growth. These need to be validated before submission to iGEM.

. A plan was made into the characterisation of PyeaR (BBa_K1001) was thought through and discussed between Lukas, Rebecca and Joy. We plan to grow up PyeaR + GFP in cultures reading the absorbance regularly (hourly). The inoculation will be made directly from the plate. The pilot study will last for 6 hours and the full study if the pilot study goes according to plan will be 12 hours. To generate a comparison of PyeaR against non transformed cells, Bioline competent cells were plated and grown for next day use.

Day 2

Labs

. LB media and Agar media was made in vast quantities.

. To produce more M-B and B-M plasmid (already ligated into pSB1C3 plasmids) DNA, more colonies were inoculated into LB media. We plan to maintain a running culture of this, whereby we inoculate this into fresh culture daily.

. Rebecca did a 9 hour growth study with PyeaR transfoormed cells and Bioline alpha E.coli cells as a comparison. This was done with wavescan function and single wavelength absorbance at 660nm which is not absorbed by pure LB. Recordings were taken every hour and a serial dilution was made and the two lowest dilutions were plated (10^-4 and 10^-5).

Day 3

Labs

. Forum preparation - plates with Biobrick written with one letter per plate (using the pYEAR with GFP and potassium nitrate to induce promoter activity)

. Rebecca, Khadija and Lucas, used day 2 samples and carried out a growth study with a reading every minute for 1 hour.

Day 4

- Forum preparation - plates used for the forum science day had their fluorescence intensified by adding more potassium nitrate to the cells on the plate.

Day 5

. UK Team Meet-Up! The team took the day off lab inorder to enjoy and host a great UK iGEM team meet up at the google campus, london (Cheak out human practices tab for more infomation about this day).