Team:Calgary/28 July 2010

From 2010.igem.org

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Today, I started the constructions of degP and CpxR promoters (K135000 and K239000) with the RFP and GFP constructs (I13507 and I13504) using the restriction enzymes XbaI, SpeI, and PstI. The constructions were done into AK plasmid backbones. As well, I started the PCR with primers that were previously designed by Alex and Patrick to take the CpxP promoter from the E. coli genome. The PCR finished but it ended too late to run on a gel. Tomorrow, that gel will be run. I continued contacting companies about possible sponsorships for our project. Our current sponsors will be posted on the website soon.
Today, I started the constructions of degP and CpxR promoters (K135000 and K239000) with the RFP and GFP constructs (I13507 and I13504) using the restriction enzymes XbaI, SpeI, and PstI. The constructions were done into AK plasmid backbones. As well, I started the PCR with primers that were previously designed by Alex and Patrick to take the CpxP promoter from the E. coli genome. The PCR finished but it ended too late to run on a gel. Tomorrow, that gel will be run. I continued contacting companies about possible sponsorships for our project. Our current sponsors will be posted on the website soon.
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<u>Himika</u>
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The plates with plasmid switch from July 27, 2010 grew. I did a PCR of 11 colonies of B1 and 11 colonies of C1. I ran a 1% agarose gel of the PCR products with the master mix as controls. The PCR unfortunately did not work. B1 DNA gave wrong sized bands but C1 gave me no bands at all.  Seems like there was no replication for the C1 colonies. I will not provide pictures of gel because it has absolutely no bands. Therefore I will reconstruct tomorrow.  I also ran a restriction digest of the different versions of I0500 (C1-D1, C1-D2, C2-D1, C2-D2) with EcoRI/PstI and SpeI/XbaI that we have sitting in our freezer to check which of the plasmids have the right size bands***. I also did research on what factors are responsible for inclusion body formation and how the mechanism of inclusion body formation differs in each case.
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Revision as of 00:56, 29 July 2010

Wednesday July 28, 2010


Patrick

The progress on learning Autodesk Maya seems to be going quite smoothly. I spent much of the day reading through the basic tutorials. I now know how to manipulate and create various polygonal and smooth-surface shapes. Tomorrow I will continue learning the program in polygonal modelling.


Chris

Today, I started the constructions of degP and CpxR promoters (K135000 and K239000) with the RFP and GFP constructs (I13507 and I13504) using the restriction enzymes XbaI, SpeI, and PstI. The constructions were done into AK plasmid backbones. As well, I started the PCR with primers that were previously designed by Alex and Patrick to take the CpxP promoter from the E. coli genome. The PCR finished but it ended too late to run on a gel. Tomorrow, that gel will be run. I continued contacting companies about possible sponsorships for our project. Our current sponsors will be posted on the website soon.

Himika

The plates with plasmid switch from July 27, 2010 grew. I did a PCR of 11 colonies of B1 and 11 colonies of C1. I ran a 1% agarose gel of the PCR products with the master mix as controls. The PCR unfortunately did not work. B1 DNA gave wrong sized bands but C1 gave me no bands at all. Seems like there was no replication for the C1 colonies. I will not provide pictures of gel because it has absolutely no bands. Therefore I will reconstruct tomorrow. I also ran a restriction digest of the different versions of I0500 (C1-D1, C1-D2, C2-D1, C2-D2) with EcoRI/PstI and SpeI/XbaI that we have sitting in our freezer to check which of the plasmids have the right size bands***. I also did research on what factors are responsible for inclusion body formation and how the mechanism of inclusion body formation differs in each case.

No notebook page exists for this date. Sorry!