Plasmid DNA
J.P. Latonio, R.E. Loquellano, N. Lustre, M.M. Manalo, E. Marasigan, and F.
Marzan
4Bio5, College of Science, University of Santo Tomas, Espaa, Manila
Summary
Keywords:
Restriction endonuclease,
pBR322, ethidium
bromide, agarose gel
electrophoresis
Introduction
Restriction
enzymes
are DNA-cutting
enzymes found in bacteria. Because they cut
within the molecule, they are often
called restriction endonucleases. A restriction
enzyme recognizes and cuts DNA only at a
particular sequence of nucleotides called its
recognition site. The rarer the site it recognizes,
the smaller the number of pieces produced by a
given restriction endonuclease. Some of the
8 control
Figure1
The results of the gel are shown in Figure 1.
The number of restriction enzymes added to the
microcentrifuge tubes corresponds to the
number of expected bands to be seen, which
then signifies the number of segments formed
after the cleavage. In the figure shown, only
lanes 6, 7 and 8 showed bands, which confirms
that different enzymes cleaved the plasmid,
although the expected results were not achieved.
Ideally, lane 6 should have two bands
representing the two segments cleaved by the
two enzymes namely Bam HI and Eco RI as
well as lane 7 which should have two bands
Figure 2