Restriction digest of phage lambda DNA and Agarose gel elektrophoresis - Restriction enzymes

10 important questions on Restriction digest of phage lambda DNA and Agarose gel elektrophoresis - Restriction enzymes

What are restriction enzymes?

It are endonucleases, i.e. can cleave double-stranded DNA at specific locations in the DNA chain, the restriction sites.

What is the function of restriction enzymes?

These enzymes recognize a specific base sequence in the DNA and hydrolyze (= cleave) only at these specific locations (recognition sequences) in the DNA the phosphodiester bonds between the nucleotides.

Where does the recognizable base sequence consist of?

At least four base pairs and is characteristic for each enzyme. Thus different restriction enzymes recognize different base sequences of possibly different size.
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When are sticky end created and what are they?

When the cleaving is not done symmetrically, these are short pieces of single-stranded DNA that stick out at the places where the restriction enzyme has cleaved.

What happens if the cleaving is symmetrically?

Blunt ends are created. These are DNA end without single-stranded DNA sequences.

At which locations does the EcoRI cleave the DNA?

Locations where the base sequence GAATTC (CTTAAG) occurs.

From which bacterium orgins the HindIII-enzyme?

Haemohilius influenzae

At which location cleaves the HindIII-enzyme?

At location with recognition sequence AAGCTT

Where does the BamHI orginates from?

From the bacterium Bacillus amylolique-faciens

Enzyme BamHI cleaces the DNA at locations where the sequences

GGATCC occurs

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