At this point in time, girls were allowed to choose their own husbands rather than have parents assign them. By her finding love, it shows extreme hope because she’ll be able to have a really happy life.


Plasmids, enzymes, and gel electrophoresis were all things used in this project. The purpose was to use all of the knowledge I have learned in a biotech class to identify an unknown plasmid which was named “antimony”. It was important that I could identify the plasmid so that I could prove that I know what I’m doing in a laboratory and so that I can also demonstrate that I know how to identify a plasmid. A plasmid is a circular piece of DNA that comes from bacteria. It can transport and copy itself. Scientists like to use them to take a desirable trait from one bacterium and put it in another. Restriction enzymes cut the plasmid in certain locations which can be very useful to scientists. Forensic scientists can use restriction enzymes to identify a killer by comparing DNA sizes and other scientists can see where DNA fragments are positioned. Gel electrophoresis separates DNA based on the size of it or the number of base pairs. DNA is put in wells in an agarose gel which has been placed in an electrode chamber with the wells on the negative end of the chamber. Since DNA is negative, when the chamber is turned on the DNA moves toward the positive side of the gel. To do so, it has to move through small pores in the gel and the smaller sizes move fastest, so the DNA ends up separated based on size. I used restriction enzymes to cut the plasmid and then used gel electrophoresis to separate the DNA. I then compared the results with predicted results of all 4 potential plasmids to discover which was mine. 
	The first thing done to begin was to decide how many digests were to be run. It was decided that 4 digests would take place. The first would be no digest at all, the second would use BglI (made by NEB), the third would use HindIII (also made by NEB) and the fourth would be a mixture of both BglI and HindIII. Using micropipettes, 11.5 µL of antimony, which was at 0.2 µg/ 11.5 µg concentration (this was given to me by my instructor), and 1.5 µL of NEBuffer 2 (obtained from NEB) were added to 4 appropriately labeled PCR tubes. In the tubes labeled BglI and B+H, 1 µL of BglI was transferred into each. HindIII was added to the B+H tube as well and was also placed in the HindIII tube. After that, dH2O was added to each tube to bring up their levels to exactly 15 µg. In the fourth tube, labeled C, only the DNA, NEBuffer2 and dH2O were added because this one was the control tube. 