| Line 10: | Line 10: | ||
. 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. | . 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. | ||
| - | |||
==Day 2== | ==Day 2== | ||
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 |
. 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.
. The pilot study was carried out for the planned 6 hours. From the plate a colony was selected and inoculated into 15ml of LB media in a sterile tube. Into this potassium nitrate was added at 0mm, 5mM and 10mM concentration. E.coli cells were inoculated into the same amount of media without the addition of potassium nitrate. Every hour an absorbance reading was taken and a serial dilution was made. Of the dilutions as we do not know the number of cells to be expected 3 of the dilutions were plated (10^-3, 4 and -5).
. LB media and Agar media was made in vast quantities and autoclaved.
. 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.
. The plates from the previous day's pilot study was viewed. It was found that initially, there was an insufficient number of cells in the culture to plate and hence within the first 3 hours, there were little to no cells on the plates. With progression of time there were more colonies.
. We decided to make the study longer and take both full wavescans as well as absorbance readings at 660nm. Instead of plating constantly . 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.
- Forum preparation - plates used for the forum science day had their fluorescence intensified by adding more potassium nitrate to the cells on the plate.
. 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).
"
