{{:Team:Northwestern/Templates/trial}}

{{:Team:Northwestern/Templates/trial}}

*Started a restriction digest and gel extraction of our miniprepped DNA. We wanted to see if we could isolate the part that we had taken from the registry and transformed.  

*Started a restriction digest and gel extraction of our miniprepped DNA. We wanted to see if we could isolate the part that we had taken from the registry and transformed.  

*Transformed all four types of competent cells that we have made so far with the same positive control. We want to try and get an idea of how competent each batch is.  

*Transformed all four types of competent cells that we have made so far with the same positive control. We want to try and get an idea of how competent each batch is.  

*Transformed a constitutive promoter and several reporter genes for use in our project

*Transformed a constitutive promoter and several reporter genes for use in our project

*Grew bacteria with the Las and Rhl related parts from the registry from our glycerol stocks overnight so that we could miniprep them and extract DNA tomorrow.  

*Grew bacteria with the Las and Rhl related parts from the registry from our glycerol stocks overnight so that we could miniprep them and extract DNA tomorrow.  

*The cell competency test was a success. All of our plates had colonies, but some had far more than others. Thus we now know to use those cells as much as possible in our transformations.  

*The cell competency test was a success. All of our plates had colonies, but some had far more than others. Thus we now know to use those cells as much as possible in our transformations.  

*The four transformations from yesterday also appeared successful, with a number of colonies appearing on each plate.  

*The four transformations from yesterday also appeared successful, with a number of colonies appearing on each plate.  

*After the miniprep, we tested the samples and found no DNA. We quickly realized the overnight cultures did not appear to contain any growth. This means either our glycerol stock was bad or something went wrong even earlier. We decided the safest thing to do was re-transform all the affected parts.  

*After the miniprep, we tested the samples and found no DNA. We quickly realized the overnight cultures did not appear to contain any growth. This means either our glycerol stock was bad or something went wrong even earlier. We decided the safest thing to do was re-transform all the affected parts.  

*Started overnight cultures with yesterday’s reporter transformations

*Started overnight cultures with yesterday’s reporter transformations

*The new transformations were successful, although some of the plates had very little colonies.  

*The new transformations were successful, although some of the plates had very little colonies.  

*The overnight cultures were cloudy, so we went ahead with our miniprep.  

*The overnight cultures were cloudy, so we went ahead with our miniprep.  

*Once digestion was complete, we began to ligate a constitutive promoter together with the LasR gene. We’re trying the 3A assembly for now, and we have the DNA from the second miniprep in waiting in case something goes wrong. However, we had to postpone this because we forgot to cut the plasmid backbone!

*Once digestion was complete, we began to ligate a constitutive promoter together with the LasR gene. We’re trying the 3A assembly for now, and we have the DNA from the second miniprep in waiting in case something goes wrong. However, we had to postpone this because we forgot to cut the plasmid backbone!

*We also ran out all of our 4 digestion products on a gel, but the bands did not seperate well.  

*We also ran out all of our 4 digestion products on a gel, but the bands did not seperate well.  

*We miniprepped the overnight cultures of the remaining five parts.  

*We miniprepped the overnight cultures of the remaining five parts.  

*We digested those 5 parts, along with DNA from yesterday’s second miniprep, and plasmid backbones

*We digested those 5 parts, along with DNA from yesterday’s second miniprep, and plasmid backbones

*We began to ligate part combinations together with the 3A assembly, and transformed them with some extra competent cells from one of our advisors. We also have a standard assembly based method prepared in case this fails.  

*We began to ligate part combinations together with the 3A assembly, and transformed them with some extra competent cells from one of our advisors. We also have a standard assembly based method prepared in case this fails.  

*We made new glycerol stocks of all our parts.

*We made new glycerol stocks of all our parts.

*After observing that our 3A assembly transformations did not work, we brainstormed possible places where errors may have occurred.  

*After observing that our 3A assembly transformations did not work, we brainstormed possible places where errors may have occurred.  

*In the meantime, we started 1A standard assembly to see if we will get better results from this method.

*In the meantime, we started 1A standard assembly to see if we will get better results from this method.