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BT 201 Biotechnology Techniques I

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1 BT 201 Biotechnology Techniques I
Restriction Enzymes BT 201 Biotechnology Techniques I

2 What Are Restriction Enzymes?
Endonucleases produced by prokaryote organisms that are capable of cutting DNA at specific sequences Found naturally in bacteria, used as a defense mechanism against invading nucleic acids Exploited by molecular biologists to cut DNA to be studied or modified at will

3 How Restriction Enzymes Work
RE’s recognize a specific base sequence in the DNA, depending on what enzyme you are using RE’s cut the DNA between the specific bases within or near that sequence, called the restriction site The result of the cut can either be “blunt” or “sticky” ends

4 Types of Restriction Enzymes
Type I: cut 100 bases after the restriction site Type II: cut at the restriction site Type III: cut bases after the restriction site The type of enzyme used depends on the desired outcome

5 Blunt Cuts Do not have an overhang
Always compatible with other blunt ends for ligation purposes Some enzymes use this type of cut

6 Sticky Cuts Leave an overhang of bases
Sequence of material to be ligated must be complementary to overhang Many enzymes use this type of cut

7 Where Do We Get Them? Commercially there are several hundred RE’s available Obtained from different species of bacteria Named for the bacteria they come from

8 Uses of Restriction Enzymes
To cut DNA (obviously) For determination of length of DNA extracted For determination of presence of insert To allow DNA to be cut and pasted For preparation of molecular standards To make a chromosome map

9 Disclaimer This workforce solution was funded by a grant awarded under the President’s Community-Based Job Training Grants as implemented by the U.S. Department of Labor’s Employment and Training Administration.  The solution was created by the grantee and does not necessarily reflect the official position of the U.S. Department of Labor.  The Department of Labor makes no guarantees, warranties, or assurances of any kind, express or implied, with respect to such information, including any information on linked sites and including, but not limited to, accuracy of the information or its completeness, timeliness, usefulness, adequacy, continued availability, or ownership.  This solution is copyrighted by the institution that created it.  Internal use by an organization and/or personal use by an individual for non-commercial purposes is permissible.  All other uses require the prior authorization of the copyright owner.

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