There are three classes of restriction enzymes; Class I, Class II and Class II. Nevertheless, Class II is most popular in recombinant technology. Explain the reason behind this.
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Q: molecular scissors", justify that
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- Cloning Genes Is a Multistep Process In cloning human DNA, why is it necessary to insert the DNA into a vector such as a bacterial plasmid?Restriction enzymes are referred to as "molecular scissors", justify thatWhy does Northern blotting not require the use of restriction enzymes? a. Restriction enzymes cut at specific sites within DNA is this right?
- Restriction sites are palindromic; that is, they read the same in the5' to 3' direction on each strand of DNA. What is the advantage ofhaving restriction sites organized this way?If restriction endonucleases are produced by bacteria within a host, why don’t these enzymes chew up the genomic DNA of their host? What is the role of DNA methyltransferase in this? Indicate the answertrue or false if a restriction enzyme recognizes the restriction site, 5' AACGTT3', and the enzyme cuts between the second A and the C, this will produce a "sticky end," which is useful for cloning into a plasmid vector.
- Why is it possible to visualize a PCR product on an agarose gel even if the template genome is present at such a low concentration that it cannot be seen?A microbiologist discovers a new type II restriction endonuclease. When DNA is digested by this enzyme, fragments that average 1,048,500 bp in length are produced. What is the most likely number of base pairs in the recognition sequence of this enzyme?A 2.0kb bacterial plasmid ‘BS1030’ is digested with the restriction endonuclease Sau3A; the plasmid map is depicted in the diagram below and the Sau3A (S) restriction sites are indicated. Which of the following DNA fragments do you expect to see on an agarose gel when you run Sau3A-digested plasmid ‘BS1030’ DNA? a. 250 bp, 450 bp, 550 bp, 1.1 kb, 1.5 kb and 2.0 kb b. 2.0kb c. 250 bp, 400 bp, 450 bp, 500 bp and 550 bp d. 100 bp, 200 bp, 250 bp, 400 bp, 500 bp and 550 bp
- You have a recombinant plasmid containing a vector and a segment of foreign DNA, both equal sizes. Draw a picture of this recombinant plasmid labeling foreign and vector regions. Where the foreign DNA meets the vector, there is a cut site for restriction enzyme ABC1. When the recombinant plasmid is cut by ABC1, how many fragments do you expect to be produced? Identify these fragments.Restriction enzymes and DNA ligase play essential roles in DNA cloning. How is it that a bacterium that produces a restriction enzyme does not cut its own DNA? Describe some general features of restriction sites.How many fragments would you expect to be formed from digestion of a 2500 base pair long linear piece of DNA using a restriction enzyme with a 5 base pair recognition sequence?”