"
Team:UC Davis/Notebook/Week 5
From 2012.igem.org
(Created page with "Monday We tried to make compotent cells for MG1655 strain, but put them in the 37 degree room instead of at room tempurature and messed that up. We liquid cultured and plated D...") |
|||
| Line 1: | Line 1: | ||
Monday | Monday | ||
| - | We tried to make compotent cells for MG1655 strain, but put them in the 37 degree room | + | Our team was finally back together, as a majority of people were absent for the latter part of last week. A few of us went over what was done last week, and how there was an error in running the gel extractions, so we made a decision to re-attach the promoters to the B0034 RBS. Some of us began by running digestions of J23100, K206000, J23101, and R0010 at the E and S sites. B0034+C0012 was digested at X and P, J23101 was digested at S and P, and B0034 was digested at E and X. We also wanted to carry on some of the themes from last years project, and began trying to mutate K206000, or the pBAD promoter, to improve its function through random mutanogenesis. Some of our team members also began preparing the E. Coli strain, numbered MG1655, by streaking the organisms on plates from the glycerol stock. After, we rehydrated MG1655 strain. We also liquid cultured and plated DH5alpha cells and tried to make compotent cells for MG1655 strain, but put them in the 37 degree room. Some of our team members began to run error prone PCR (PROTOCOL) on the K206000 part as well, to make a part family as well. In our discussion of how we were going to assay our samples, we also ordered pNPB, another sample used to determine degredation in comparison to PET. |
| - | + | ||
Tuesday | Tuesday | ||
| - | + | When the team came into lab today, we used the spectrophotometer and the OD readings were well over 1, which signifies we overgrew them. The error came from placing the cells in the 37 degree room instead of at room tempurature. Oops. So we then remade the component cells for the MG1655 strain and extracted genomic DNA from the MG1655 and DH5alpha E. Coli strain. | |
Revision as of 21:46, 20 July 2012
Monday
Our team was finally back together, as a majority of people were absent for the latter part of last week. A few of us went over what was done last week, and how there was an error in running the gel extractions, so we made a decision to re-attach the promoters to the B0034 RBS. Some of us began by running digestions of J23100, K206000, J23101, and R0010 at the E and S sites. B0034+C0012 was digested at X and P, J23101 was digested at S and P, and B0034 was digested at E and X. We also wanted to carry on some of the themes from last years project, and began trying to mutate K206000, or the pBAD promoter, to improve its function through random mutanogenesis. Some of our team members also began preparing the E. Coli strain, numbered MG1655, by streaking the organisms on plates from the glycerol stock. After, we rehydrated MG1655 strain. We also liquid cultured and plated DH5alpha cells and tried to make compotent cells for MG1655 strain, but put them in the 37 degree room. Some of our team members began to run error prone PCR (PROTOCOL) on the K206000 part as well, to make a part family as well. In our discussion of how we were going to assay our samples, we also ordered pNPB, another sample used to determine degredation in comparison to PET.
Tuesday
When the team came into lab today, we used the spectrophotometer and the OD readings were well over 1, which signifies we overgrew them. The error came from placing the cells in the 37 degree room instead of at room tempurature. Oops. So we then remade the component cells for the MG1655 strain and extracted genomic DNA from the MG1655 and DH5alpha E. Coli strain.
