Restriction Enzyme

 Introduction    

    We started this week off with restriction enzymes. Learning about restriction enzymes is imperative to developing laboratory methods as well as learning about genetic engineering. A restricted enzyme is a protein that has been isolated from bacteria, producing DNA fragments. We started with 2 micro-liters of restricted enzymes, 10 micro-liters of our buffer, 8 micro-liters of our kanamycin DNA, and then 80 micro-liters of PCR water. The main goal was to remove pst1 from our sample's DNA. 




After we had our samples combined, we incubated at 37 degrees celsius for 15 minutes, which was then followed by inactive at 80 degrees celsius for 20 minutes. We then loaded the samples into our gels and then ran the gels at 85 mA for around 30 minutes. 


Once the gel had run its course, we placed the gel over the UV light and found out the kanamycin did not split how we were hoping it would. 


The following day we ran an additional gel, but this time it was a 2% gel, which consists of 30 mL of 1x TAE and 600 mg of agarose powder. Similar to the first run, we did not get the results we wished for. 




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