PCR

 Introduction

    This week we hit the ground running. We started by taking our fragments of DNA and started to put them together. We took 3 different samples, two being DNA and the other being our plasmid. Inside of the samples we had 1 micro-liter of our forward primer, 1 micro-liter of our reverse primer, 2 micro-liters of the DNA/Plasmid, and finally 16 micro-liters of master mix. Once all the samples were combined, they were placed inside the thermal cycler to run their course. Upon completion of the thermal-cycler, we came to find out there was a mix-up in which DNA/Plasmid was placed into our tetramycin sample. Since we took the time to create the samples and had them run in the thermal cycler, we decided we would run the sample on the nano-drop as well as run a gel. 


    As seen from the gel there are a couple ghost fragments as well as blurriness, we'll be working towards cleaning the sample for the cleanest fragments we can get. 

    The following time we met in the lab the sampling issue had been corrected on our left and right fragments and they were to be placed in a bigger gel (consisting of 120 mL 1x TAE and 1200 mg Agarose powder).

    In addition to the current project we had been working on, we also tested the growth of deinococcus radiodurans in the Mueller-Hinton medium. Since ASU has specific parameters regarding testing, we had to grow D-RAD in the MH medium specifically for them. We took our sample to the nano-drop to confirm whether the cells were able to live in the medium, which they did grow.  

    At the end of the day, we began to remake tetramycin samples with the correct fragments. Same as the previous samples we did 16 micro-liters of master mix, 2 micro-liters templates, 1 micro-liter plasmid-5, and 1 micro-liter plasmid-6. The samples were then placed back inside the thermal-cycler on a gradient (different temperatures per row) to find the optimum fostering temperature. 



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