"
Week19 10/17/10-10/23/10
From 2010.igem.org
Week19 Highlights
We retried assembly of CP-LacPI both phosphatase treated and non-phosphatase treated. We were unable to assemble a full CP-LacPi-GFP part. We submitted 3 of our DNA constructs to the parts registry.
10/17/10
Timi phosphotase-treated the newly digested chlor backbone and ligated a second set of CP-LacPI. Kevin transformed and plated this new set of phosophotase treated Cp-LacPI.
Kevin inoculated the first set of non-phosphotase treated CP-LacPI inoculations.
10/18/10
Matt miniprepped 31GFP, 21GFP (old), and 32GFP (old). He digested the miniprepped DNA with E/P for Kevin to run on a gel. He also reinoculated all 9 CP-LacPI combinations.
Matt inoculated the phosophotase-treated ligations at 9pm.
10/19/10
Plate reader with Kevin @ 3pm.
Sean - ran CHS3(pMAL+SDM) digest, pMAL digest, and ligation on a gel -> CHS3 exhibited incorrect bands (there are 2, when there should be 1) ligation showed nothing -> plan to run CHS3 undigested and rerun the above 3.
10/20/10
Timi and Kevin ligated the old 212, 312, and 322 parts into a phosophotase-treated Chlor backbone for submission into the registry. They transformed and plated for inoculation tomorrow night.
They inoculated cultures for the plate reader.
10/21/10
Timi/Kevin: Early morning plate reader protocol entering.
Weekly lab meeting with advisors.
10/22/10
PCR'ed out CHS3 with pMAL ends and Gel extracted
Kevin was in lab until 3am to see if any of the CP-LacPI parts were ligated to GFP. He ran multiple gels and sent the results to Prof. Leonard. Unfortunately, we do not have the construct ready. Thus, we will likely not be able to characterize a CP-LacPI part by Wednesday. However, we plan on working to characterize one by the Jamboree.
10/23/10
Timi miniprepped the overnights for DNA that will be submitted the registry. We will be sending in parts http://partsregistry.org/Part:BBa_K418003 BBa_K418003, http://partsregistry.org/Part:BBa_K418005 BBa_K418005 and http://partsregistry.org/Part:BBa_K418006 BBa_K418006 on Monday afternoon via Fedex.
Timi and Kevin retransformed the confirmed CP-LacPI parts (CP2-LacPI1, CP3-LacPI1 and CP3-LacPI2) from earlier in the summer into ChiA competent cells. The cells were plated onto Chlor plates. We will be continuing our work to hook them up to GFP over the subsequent weeks. Hopefully, we will have all of our work done by the Jamboree.
