Restriction enzymes are useful for many different applications. Because the DNA sequence is different in each organism, the pattern of restriction sites will also be different. … If DNA from a different organism is cut by the same restriction enzyme, a different set of fragments will be generated.
Why would different restriction enzymes cut the same DNA molecule?
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Why would different restriction enzymes generate different DNA fragments if they are all cutting the same DNA molecules?
Explanation: Restriction enzymes cut at specific sequences so the same restriction enzyme must be used because it will produce fragments with the same complementary sticky ends, making it possible for bonds to form between them.
Do all restriction enzymes cut DNA at the same recognition?
Each restriction enzyme recognizes just one or a few restriction sites. When it finds its target sequence, a restriction enzyme will make a double-stranded cut in the DNA molecule.
Can you use more than one restriction enzyme to cut DNA?
To be able to clone a DNA insert into a cloning or expression vector, both have to be treated with two restriction enzymes that create compatible ends.
Why do we use two different restriction enzymes?
The use of 2 different enzymes makes self ligation of the vector impossible and makes the insertion unidirectional. Whereas in the case of single digest, selfligation occurs and insertion may occur in both ways. Overall the use of 2 RE increases the probability to get the right construct.
What would happen if we used different restriction enzymes to cut the plasmid and the gene?
Restriction endonuclease identifies and cuts the same pallindromic sequence in both DNA and Vector due to which when they will be mixed , their complementary bases will join and it will form the r-DNA, If both are cut with different RE , then on mixing they wont ligate with each other as their bases will not match.
Why do restriction enzymes not cut bacterial DNA?
A bacterium is immune to its own restriction enzymes, even if it has the target sequences ordinarily targeted by them. This is because the bacterial restriction sites are highly methylated, making them unrecognizable to the restriction enzyme.
What do restriction enzymes cut?
Restriction enzymes cut DNA bonds between 3′ OH of one nucleotide and 5′ phosphate of the next one at the specific restriction site. Adding methyl groups to certain bases at the recognition sites on the bacterial DNA blocks the restriction enzyme to bind and protects the bacterial DNA from being cut by themselves.