Restriction Digest Involves The Uses Of Restriction Enzymes

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Restriction Analysis
Hayley Keller
Biochemistry Laboratory: BIOL 2324
TA: Manasa Madasu
Date Performed: July 14th, 2015
Date Due: July 21st, 2015 Introduction:

Restriction digest involves the use of restriction enzymes (also known as restriction endonucleases) to locate specific base pair sequences in DNA. These enzymes cut, or cleave, DNA only at their designated sequence, which is referred to as a recognition sequence. While there are four different types of restriction enzymes (1), the only type that was worked with in the following experiment were type II restriction enzymes (2). These enzymes have recognition sites that are mostly palindromic and usually consist of around four to eight base pairs. They also require only magnesium (Mg2+) as a cofactor to operate. Cofactors are molecules that bind to enzymes in order to activate them (3). Additionally, they cut DNA only at, or very near to, their specified restriction site, unlike other types, which cleave at various distances from their recognition site (1). The restriction enzymes that will be used in following experiment are Hind III, PVU II, and Bgl I (2). Hind III recognizes and cuts DNA at the sequence AAGCTT. It is isolated from Haemophilus influenzae (4), which is a bacteria that is the cause of several diseases, including pneumonia, and meningitis. (5) When Hind III is used to cleave DNA, the end result will have “sticky ends,” which means that there will be a few unpaired nucleotide bases on each end of
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