Team:Calgary/9 June 2010


Wednesday June 9, 2010

Alex's and Patrick's streak plates. Photograph taken at 9:50am June 9th, 2010.


Today I Miniprepped the overnight cultures of B0034 and B0015 which were made yesterday. A colony PCR of the B0034-E1010 construct was done as well in order to verify that the construction worked.


Today I ran my restriction digest of I0500 and I13453 on a gel. The band sizes were not quite what was exepcted. The I13453 was not cut at all as there was only 1 single band in each lane. I repeated this digest overnight and the results will be visualized tomorrow. The I0500 was cut, however Colony 1 showed bands much larger than expected so I will proceed with colony 2. I did a construction digest of I0500 C2 with B0034 and then transformed into competant TOP10 cells and plated for overnight growth. Today I also spent some time cleaning out the freezer and labelling boxes so that we can start to organize our constructs.

Dev, Chris, and Jeremy

Today, they Miniprepped the plasmid backbones pSB1A2, pSB1AC3, pSB1AK3, and pSB1K3 so they could be used in a plasmid transfer. The resulting plasmid of pSB1K3 was then constructed to contain the parts of E0032 and E0040. The plasmid transfer was done so when the parts of E0032 and E0040 (fluorescence proteins) are constructed with promoters and RBS, the antibiotic resistances would be different.

Alex and Patrick

The streak plates were successfully done, though the colonies appear to be a little close together. We'll have to be careful if we need to take another colony. We miniprepped the overnight cultures and harvested the plasmids. The analyzed data is also here.

Using these samples, we're trying to clone the R0040 promoter in front of the E0430 part in order to test the effectiveness of the EYFP present in E0430. We're only using the two best concentrations of each part to construct a (hopefully) functioning device. We will have to transform them tomorrow.

We're also trying out using NEBuffer 2 and NEB SpeI enzyme since our lab ran out of Invitrogen REact 1 buffer, which would have been our first choice. Also, we used T4 Ligase and T4 Ligase buffer and left the reaction at room temperature overnight because we did not have time to transform them today. We can leave the plasmid in this ligation state until tomorrow.