What is restriction enzyme digestion of DNA?

Restriction Digestion is the process of cutting DNA molecules into smaller pieces with special enzymes called Restriction Endonucleases (sometimes just called Restriction Enzymes or RE's).

What is the purpose of restriction digestion of plasmid DNA?

Restriction digestion of recombinant plasmid constructs provides a fast, cost-efficient method of gaining indirect sequence information. Multiple plasmid constructs can be analyzed simultaneously for the presence or absence of an insert, orientation of the insert, plasmid size, and some site-specific sequence data.

What are two functions of restriction enzymes?

The function of restriction endonucleases is mainly protection against foreign genetic material especially against bacteriophage DNA. The other functions attributed to these enzymes are recombination and transposition.

What is the purpose of restriction enzymes in PCR?

Restriction enzymes can also be used to generate compatible ends on PCR products. In all cases, one or more restriction enzymes are used to digest the DNA resulting in either non-directional or directional insertion into the compatible plasmid.

Why is it important to perform a restriction digest after cloning?

Digestion Set up restriction digests for your insert (or donor plasmid) and plasmid backbone. Because you lose some DNA during the gel purification step, it is important to digest plenty of starting material.

How is restriction enzyme digestion set up?

Set up the reaction using the following scheme: 1) Determine the amount (total ug and total ul) of DNA to be digested. 2) Use the ug amount of DNA to determine how many enzyme units to use. 3) Determine how many ul of enzyme to use, using the enzyme concentration. 4) Choose a total volume for the reaction.

What is the main purpose of using restriction enzymes in restriction enzyme digestion quizlet?

What is the natural function of a restriction enzyme? Their natural function is to destroy foreign DNA entering the cell by cleaving the bacteriophage DNA to prevent infection. The cell's own DNA is modified by methylation to protect it from its own enzyme. Each restriction enzyme has a specific methylase.