What do restriction enzyme HaeIII do?
HaeIII is one of many restriction enzymes (endonucleases) a type of prokaryotic DNA that protects organisms from unknown, foreign DNA. It is a restriction enzyme used in molecular biology laboratories. It was the third endonuclease to be isolated from the Haemophilus aegyptius bacteria.
Which allele will HaeIII cut?
dominant allele
Restriction enzymes cut at specific DNA sequences termed recognition sequences. HaeIII recognizes a 5′ GGCC 3′ DNA sequence (refer to Figure 1, page 2). The dominant allele contains the 5′ GGCC 3′ recognition sequence and therefore will be cut by HaeIII.
How many DNA fragments are produced by HaeIII?
Thus treatment of this DNA with the enzyme produces 11 fragments, each with a precise length and nucleotide sequence. These fragments can be separated from one another and the sequence of each determined. HaeIII and AluI cut straight across the double helix producing “blunt” ends.
What does HaeIII recognize?
This entry represents the restriction endonuclease HaeIII, which recognises and cleaves the double-stranded sequence GG^CC.
What is the restriction site sequence cut by HaeIII?
The restriction enzyme HaeIII recognizes and cuts the DNA sequence 5′-GGCC-3′ 3′-CCGG-5′ A.
What sequence does the restriction enzyme HaeIII recognize?
Hae III recognizes the sequence GG/CC and generates fragments with blunt ends (1). Compatible ends Hae III generates compatible ends to any blunt end.
What DNA sequence does the restriction enzyme HaeIII recognize?
HaeIII recognizes a sequence that includes GGCC and cleaves between the G and C, leaving a blunt-ended fragment. To determine the cleavage site of CbeI, a DNA fragment containing three HaeIII recognition/cleavage sites (Fig. 3a) was used as substrate with purified CbeI enzyme.
Why is HaeIII added to the PCR product?
Subsequently, the purification of the PCR product will be carried out and it will be digested with the restriction enzyme HaeIII to identify the presence or absence of the SNP linked to the phenotype of detecting the bitter taste in PTC papers.
What enzyme cuts this DNA sequence 5?
restriction enzyme, also called restriction endonuclease, a protein produced by bacteria that cleaves DNA at specific sites along the molecule.
How many fragments are produced by BamHI?
Under ideal conditions there would be 6 fragments from Enzymes A and B, and 8 fragments from Enzyme C. GGATCC is the recognition site for BamHI and is found in λ DNA at 5 locations.
What type of enzyme cuts DNA and leaves sticky ends?
A restriction enzyme is a DNA-cutting enzyme that recognizes specific sites in DNA. Many restriction enzymes make staggered cuts at or near their recognition sites, producing ends with a single-stranded overhang.
What is the HaeIII methyltransferase gene?
HaeIII Methyltransferase modifies the internal cytosine residue (C5) of the sequence GGCC. This enzyme comes from an E. coli strain that carries the cloned HaeIII modification gene from Haemophilus aegyptius (ATCC 11116).
How do you incubate HaeIII methyltransferase?
HaeIII Methyltransferase is incubated with 1 µg of λ DNA in 10 µl of 1X HaeIII Methyltransferase Reaction Buffer, supplemented with 80 µM S-adenosylmethionine, for one hour at 37°C followed by 15 minutes at 65°C. The extent of protection is determined by the addition of 40 µl NEBuffer 2.1 and 10 units of HaeIII restriction endonuclease.
Where does the HaeIII enzyme come from?
This enzyme comes from an E. coli strain that carries the cloned HaeIII modification gene from Haemophilus aegyptius (ATCC 11116). An E. coli strain that carries the cloned HaeIII modification gene from Haemophilus aegyptius (ATCC 11116).