Scribd
Upload
746 views

Restriction Mapping Sample Question and Its Solution

This document provides information to construct a restriction map of a circular plasmid that was digested with PstI, HindIII, and EcoRI restriction enzymes. The fragment sizes from each digest are given, as well as the fragment sizes from double digests. Using this information, the document guides drawing the restriction map, placing each enzyme site and noting the distance between sites.

Uploaded by

Aditya Lohia
Copyright
© Attribution Non-Commercial (BY-NC)
Available Formats
Download as DOC, PDF, TXT or read online on Scribd
746 views

Restriction Mapping Sample Question and Its Solution

This document provides information to construct a restriction map of a circular plasmid that was digested with PstI, HindIII, and EcoRI restriction enzymes. The fragment sizes from each digest are given, as well as the fragment sizes from double digests. Using this information, the document guides drawing the restriction map, placing each enzyme site and noting the distance between sites.

Uploaded by

Aditya Lohia
Copyright
© Attribution Non-Commercial (BY-NC)
Available Formats
Download as DOC, PDF, TXT or read online on Scribd
You are on page 1/ 1

Q.

You digest a circluar plasmid with PstI, HindIII, and EcoRI and obtain the following fragment sizes (in kb) from each of the digests shown below. Draw the restriction map this plasmid. Include the distance between each restriction enzyme site in your map. Here are the pts. PstI 6.8, 5.9 HindIII 6.5, 6.2 EcoRI 9.2, 3.5 PSTI + HindIII 4.8, 4.2, 2.0, 1.7 PstI + EcoRI 5.4, 3.8, 3.0, 0.5 EcoRI + HindIII 6.2, 3.5, 1.8, 1.2 Ans. You'll start by drawing a small circle inside of a larger circle (use pencil and be prepared to redraw, it's a bit messy to make one of these by hand). 2) At the top draw a straight line between the circles (like on the image link above). This line represents "zero" or a starting point. 3) Each cut represents the presence of 1 restriction enzyme sites. For example, PstI makes 6.8 and 5.9 fragments. That means that the two sites are located almost completely opposite of each other. There are 6.8 kilo bases (6,800 bases) between the sites on one side and 5.9 between them on the other. Draw those into your map. 4) Next look at HindIII. There are two sites located opposite from one another with 6.5 and 6.2 between them (note that when you add these up, it comes to 12.7... when you add up the PstI fragments, they also total 12.7, so the total plasmid is 12.7 kbp). Draw two HindIII sites between the PstI sites (one on each side). 5) Now look at the HindIII/PstI fragments and write in the fragment lengths between the PstI and HindIII sites. You don't need the space to scale correctly, just put the numbers wherever for now. 6) Add the EcoRI sites next. You know there are two EcoRI sites because EcoRI digestion makes only 2 fragments. You also know that the sites are somewhat close together (not opposite ends like PstI and HindIII) because one fragment is very short compared to the other. So place the EcoRI sites 6.2, 3.5, 1.8, and 1.2 away from HindIII fragments. 7) Double check the distances between PstI and EcoRI sites.

You might also like