A couple of questions

Why do we most commonly used type 2 restriction endonucleases in cloning as opposed to the other types of restriction endonucleases?

Why are Type 2 restriction enzymes used for cloning?

Type II restriction enzymes are the familiar ones used for everyday molecular biology applications such as gene cloning and DNA fragmentation and analysis. These enzymes cleave DNA at fixed positions with respect to their recognition sequence, creating reproducible fragments and distinct gel electrophoresis patterns.

Why are Type 2 restriction endonucleases most useful in recombinant DNA technology?

Type II restriction enzymes have two properties useful in recombinant DNA technology. First, they cut DNA into fragments of a size suitable for cloning. Second, many restriction enzymes make staggered cuts generating single-stranded ends conducive to the formation of recombinant DNA.

Why are type II restriction endonucleases used for DNA cloning and not types I and III?

Why are type II restriction endonucleases used for DNA cloning and not types I and III? Type II restriction endonucleases cut DNA at a specific site, mostly within the recognition site, hence they are used for cloning.

What is the advantage of using 2 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.

What is Type 2 restriction enzyme?

Type II restriction endonucleases are components of restriction modification systems that protect bacteria and archaea against invading foreign DNA. Most are homodimeric or tetrameric enzymes that cleave DNA at defined sites of 4-8 bp in length and require Mg2+ ions for catalysis.

How do Type 2 restriction enzymes work?

In Type IIP restriction enzymes, the amino acids that catalyze cleavage and those that recognize the DNA are integrated into a single protein domain that cannot be effectively sub-divided. … They cleave outside of this sequence, within one to two turns of the DNA.

Why are restriction enzymes palindromic 2?

Explanation: Enzymes such as restriction enzymes have to recognize a very specific sequence in order to carry out its task. … DNA is double stranded, so it has 'two sides' to which the enzyme can bind. A palindromic sequence is the same backwards and forwards on both sides (see image below).