A couple of questions

Why is type 2 restriction enzyme used?

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.

What is type2 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.

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.

How do Type 2 restriction enzymes cut DNA?

Type II enzymes cut DNA at defined positions close to or within their recognition sequences. They produce discrete restriction fragments and distinct gel banding patterns, and they are the predominant class used in the laboratory for routine DNA analysis and gene cloning.

What are the differences between type I and type II restriction enzymes?

Type I restriction enzyme possesses a cleaving site which is away from the recognition site. Type II restriction enzymes cleave within the recognition site itself or at a closer distance to it. This is the key difference between Type I and Type II restriction enzyme.

How does a Type 2 restriction enzyme generate blunt ends?

These enzymes cleave the phosphodiester bond of double helix DNA. It can either cleave at the center of both strands to yield a blunt end, or at a staggered position leaving overhangs called sticky ends. These are the most commonly available and used restriction enzymes.

Is EcoRI a type II restriction enzyme?

Type IIP enzymes specific for 6-8 bp sequences mainly act as homodimers, composed of two identical protein chains that associate with each other in opposite orientations (Examples: EcoRI, HindIII, BamHI, NotI, PacI.) Each protein subunit binds roughly one-half of the recognition sequence and cleaves one DNA strand.

Why are the restriction sites of Type II enzymes usually palindromic?

Explanation: Enzymes such as restriction enzymes have to recognize a very specific sequence in order to carry out its task. It binds to the DNA only in one specific configuration. … A palindromic sequence also increases the chance that both strands of DNA are cut.