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Exonuclease enzymes cleave nucleotides from the ends of DNA molecules.

Endonuclease enzymes cleave a phosphodiester bond somewhere within the DNA molecule (not at the ends).

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Describe the difference between a single digest and a double digest?

Restriction enzymes (endonucleases) are used for a variety of reasons in molecular genetics, including obtaining a "map" and cloning DNA. Single digests consitute DNA being treated with one restriction endonuclease, whereas double digests contain 2 enzymes. At times, it is difficult (or not possible) to perform double digests ... especially when the 2 enzymes have very different requirements for their activities (e.g. salt concentration, temperature optimums, ...). If a DNA restriction map is known for a particular enzyme, and if the DNA is treated with this enzyme, then one can ascertain whether the digest was complete or not. However, if a restrictioin map is just being compiled, and if the DNA is treated with 2 enzymes in a double digest, at times difficulties may arise in determining the map if either (or both) enzymes did not completely digest the DNA.


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Related Questions

What is the difference between a restriction enzyme and an endonuclease?

A restriction enzyme is a type of endonuclease. Endonucleases are enzymes that cut DNA at specific sequences, while restriction enzymes specifically cut DNA at recognition sites called restriction sites.


What are the differences between endonucleases and restriction enzymes in terms of their functions and mechanisms of action?

Endonucleases are enzymes that cut DNA at specific sites, while restriction enzymes are a type of endonuclease that specifically recognize and cut DNA at specific sequences called restriction sites. Endonucleases can have various functions in DNA repair and replication, while restriction enzymes are primarily used by bacteria as a defense mechanism against foreign DNA. Both enzymes work by breaking the phosphodiester bonds in the DNA backbone, but restriction enzymes have a more specific recognition and cutting mechanism compared to other endonucleases.


What is the difference between endonuclease and exonuclease activity in DNA degradation processes?

Endonuclease activity involves cutting DNA internally, while exonuclease activity involves cutting DNA from the ends. In DNA degradation processes, endonucleases break DNA strands at specific points, while exonucleases remove nucleotides from the ends of DNA strands.


What cuts DNA at a specific sequence of nucleotides?

Restriction enzyme cut the DNA at the specific site. Xho I is an example for restriction endonuclease which cut between C and T in the sequence of -CTCGAG- at the both strands. This is highly specific and hence they are used in DNA or gene cloning.


What is the difference between an endonuclease and an exonuclease?

An endonuclease cleaves nucleic acids internally at specific recognition sites, while an exonuclease cleaves nucleic acids at the ends by removing nucleotides one at a time. Endonucleases are involved in processes like DNA repair and recombination, while exonucleases are important for proofreading during DNA replication.


What are restriction enzymes?

Restriction enzymes (also known as restriction endonucleases) are proteins which cut DNA up at specific sequences in the genome. For example, the commonly used restriction endonuclease EcoRI recognizes every point in DNA with the sequence GAATTC, and cuts at the point between the Guanine and Adenine. Interestingly, the recognition sequences for most restriction endonucleases are genetic palindromes, e.g., the sequence reads exactly the same backwards on the complementary strand. In the case of EcoRI, the two complementary DNA strands for the recognition sequence are: 5'--GAATTC ---3'3'--CTTAAG--5'


How the hydrogen bonds break when the restriction endonuclease enzyme acts on the sugar - phosphate backbone of a DNA?

Restriction endonucleases break hydrogen bonds between complementary base pairs in DNA, not the hydrogen bonds in the sugar-phosphate backbone. These enzymes recognize and bind to specific DNA sequences, then cleave the phosphodiester bonds in the backbone at specific locations, resulting in DNA fragmentation.


How do restriction sites and a restriction map relate?

Restriction sites are specific DNA sequences recognized and cleaved by restriction enzymes, while a restriction map shows the locations of these sites on a DNA molecule. A restriction map provides information on the order and spacing of restriction sites along a DNA sequence, helping to identify the size and organization of DNA fragments generated by restriction enzyme cleavage.


What are isochizomers?

Isochizomers are such restriction endonucleases which have the same recognition sequence but may have different recognition site. examples are XmaI and SmaI which have same recognition sequence 5'CCCGGG3' but SmaI cuts between C and G and XmaI cuts between first C and second C.


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How are restriction maps used?

They are used to show the lengths of DNA fragments between restriction sites in a strand of DNA.


Why is it important to use the same restriction enzyme for both cells in recombinant DNA?

Restriction enzymes are endonucleases that digest the DNA at a sequence specific site. Hind III for example cut between two As in the sequence AAGCTT in the both strand forming a sticky end. If you use this enzyme to cut in your vector DNA, you have to use the same enzyme in the insert DNA so as they can ligate by DNA ligation. This is the important use of same restriction enzyme in cloning.