What is the recognition sequence for HindIII?
Under the standard reaction conditions, the restriction endonuclease HindIII cleaves double-stranded DNA, within the recognition sequence–A/AGCTT–at the position indicated by the arrow. In the presence of dimethyl sulfoxide the substrate specificity of this enzyme is reduced and cleavages occur at additional sites.
Are the base sequences of the HindIII and EcoRI tails complementary?
6. The two tails are 5′ AATT 3′ (EcoRI) and 5′ AGCT 3′ (HindIII). They are not complementary.
What is the recognition sequence for EcoRI?
EcoRI recognizes the sequence GAATTC, and cuts both DNA strands between the G and the A nucleotides. Protruding from the cut ends will be single-stranded DNA “tails” having the sequences AATT.
What is the difference between HindIII and EcoRI?
The key difference between EcoRI and HindIII restriction enzymes is that EcoRI is a type II restriction enzyme that is isolated from E. coli, while HindIII is a type II restriction enzyme that is isolated from Haemophilus influenza. These restriction enzymes are found in bacteria and archaea.
What is the base sequence of the tail of HindIII cut DNA?
The DNA should be cut so that 5′ AGCT protrudes from each end. The ends are sticky. 6. The two tails are (EcoRI) 5′ AATT 3′ and (HindIII) 5′ AGCT 3′.
What would be the lengths of the products of a digestion with the three enzymes HindIII ApaI and PvuI?
What would be the products of a digestion with HindIII, ApaI and PvuI? The products of a digestion with HindIII, Apal, and Pvul are 657, 2,300, and 2,881 base pairs. Use the information provided in the plasmid map of the DNA scissors document to answer this question.
How is lambda DNA/EcoRI+HindIII marker made?
Lambda DNA is digested to completion with the appropriate Thermo Scientific restriction enzyme (s) and purified and dissolved in storage buffer. DNA fragments contain various sticky ends depending on the restriction enzyme used for preparation of the marker. Lambda DNA/EcoRI+HindIII Marker can be labeled radioactively with T4 Polynucleotide Kinase.
What is the difference between CLA (C) and Eco RI (E)?
The molecule cut with Cla l (C) is the most bent whereas the molecule cut with Eco RI (E) is the straightest and migrates fastest in the gel. (C) The relative mobility is plotted as a function of the base pair at the position of cutting.
How was BIOB cloned from Kohara phage?
The bioB gene was cloned by PCR from Kohara phage 3D4 containing the bio operon and incorporated between the NdeI and HindIII sites in pET23b. The resulting plasmid pJJ07 was transformed into E. coli BL21 (DE3)pLysS ( Ugulava et al., 2000 ).