Restriction Mapping

Introduction: The goal of this lab to see and compare the DNA sequence strand to three others. The original strand will be placed next to the other strands and compare and see what sites are close and related. We used three restriction enzymes Pst1, Hpa1, and Ssp1. These were the different restriction enzyme digesters. Then after setting up the DNA it is key and essential to store it in the electrophoresis chamber over night to get the results and look over. 

Purpose: The purpose of the restriction mapping of plasmid lab is to know that a restriction map of a piece of DNA is like a fragment of the DNA. The importance of knowing the number of cuts sites present in DNA sequence for each restriction enzyme, but the needed positions of those cut sites relative to another. These steps make it easier to determine similar and unfamiliar DNA sequence strands. 

Methods: 

In this lab, we had 3 different restriction enzymes in different combinations, and 5 identical DNA samples. We were provided with 5 different tubes that contained the combinations. We then took very small pipettes and placed the samples into the gell's wells. These are the specific combinations of what was placed in each well.

1. Marker Lane

2. BLANK

3. DNA & Pst1
4. DNA & Pst1/Hpa1
5. DNA & Pst1/Ssp1
6. DNA & Pst1/Hpa1/Ssp1

Once the mixtures were placed in the wells, we put the gel tray into the electrophoresis chamber. Once in the chamber, an electricity is run through all of the gels to move the DNA through the gel. The smaller pieces moved farther than the bigger pieces that stay closer to the wells. After the process was finished, we were able to put our gel on a "light-box" so we could see the bands and measure the distance of the pieces movement and compared them to the marker lane. Pictures of the gel on the light box are below.

Data:

This picture shows the distances of the marker lane as they compare to the other lanes. 

Discussion: The restriction mapping of plasmid DNA gave us near perfect results. We had a variation of strands and cutting sites. When looking at our gel tray on the light box, you can see the relationship between the sites. For an example the last strand has the same distance of cutting site. It is difficult to see the multiply cutting sites on the fourth row. This shows how the relationship between each strand of DNA differs for each other. We consider the consistencies of our trends to be very similar. No one strand is far different. Yes, there will be different cut sites, but nothing was drastically abnormal. The results from this lab were great. Anyone looking at the results could tell the correlation between each one. The results came as we predicted and how they should actually look. Again the whole of Lab Group 2 views this restriction mapping of plasmid DNA lab as a complete success, and as one of our better resulting labs. 

Conclusion: The restriction mapping of plasmid lab was a success. As the results were close to perfect, we got everything we thought we would see, and it had great variation. The lab was so successful due to the fact that we followed the instructions to the T.  When we examined the stained gel over the light it made the cut sites clear as possible. We consider this lab VERY successful. 

Reference: 

● "Circular Restriction Mapping." Circular Restriction Mapping.N.p., n.d. Web. 18 Mar. 2015. 

● "Restriction Map." Wikipedia.Wikimedia Foundation, n.d. Web. 18 Mar. 2015. 

● "Restriction Mapping." Restriction Mapping.N.p., n.d. Web. 18 Mar. 2015.