Tuesday August 10, 2010

Jeremy

Today I ligated the products of pRFP that I had done a restriction digest and antarctic phosphatase on. And also transformed and plated them. I also did a gradient PCR with the remaining amount of pRFP and am doing a colony PCR of the plates where pRFP was gained from as well as the growth from one of the plates from a previous plasmid switch. I ran all the PCR products on a gel and used the information from the colony PCR to do overnights as well as PCR purify the gradient PCR.

Emily

Today I miniprepped all of my malE and malE31 cultures from yesterday. I then set them up to be digested with the remaining enzymes that they were not cut with. If these genes are indeed biobricked, then we would expect to see, when run on an agarose gel, 1 band at ~1200 base paors for the insert, as well as one band at ~ 3kb for the psB1AC3 and psB1AK3 vectors. I ran a 1% gel of my two gradient PCR's from last night, however I got no amplification for either of them. For some reason, our primers are not able to amplify malE31SSdel. Raida proceeded by setting up a PCR reaction with PFU instead of Taq in order to try to get our stuff cloned into TOPO TA vectors. I also ran a gel of the colony PCR that Raida set up last nigth in order to verify our hypothetical I0500-I13504 and I0500-I13507 constructs. This was using the BBK-CP primers. I also helped Chris make TOP10 Competant Cells.