Where does the restriction enzyme HindIII cut?
Thermo Scientific HindIII restriction enzyme recognizes A^AGCTT sites and cuts best at 37°C in R buffer. See Reaction Conditions for Restriction Enzymes for a table of enzyme activity, conditions for double digestion, and heat inactivation for this and other restriction enzymes.
How do you determine the size of restriction fragments?
First, work out the frequency of occurrence of the restriction site as 1-in-x bases, as explained in the example for the Intermediate level calculation. Then take the size of the DNA in kb (kilobases) and multiply by 1000 to get the size in bases. Divide this by x and round to the nearest whole number.
How many fragments did restriction enzyme make in HindIII?
The HindIII digest of lambda DNA (cI857ind1 Sam 7) yields 8 fragments suitable for use as molecular weight standards for agarose gel electrophoresis (1).
What is common between EcoRI and HindIII?
EcoRI and HindIII both contain the PD.. D/EXK amino acid sequence motif. Both restriction enzymes perform very specific cleaving of the DNA. These restriction enzymes need Mg2+ as a cofactor for their specific activity.
What are characteristics of restriction sites?
A characteristic of many of these cleavage or restriction sites is their double rotational symmetry. Generally, the cleavage sites are symmetrically positioned, or palindromic. Restriction enzymes can create fragments with sticky ends, as is the case with the enzyme BamHI, or blunt ends, as with HaeIII (Table 8.1).
How many fragments did the restriction enzyme EcoRI produce?
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.
Is pvu 1 a restriction site?
PvuI is a restriction endonuclease that is used for molecular biological applications to cleave DNA at the recognition sequence 5′-CGAT/CG-3′ to generate fragments with 3′-cohesive termini.
What is the restriction site of HindIII?
What is a HindIII site?
HindIII (pronounced “Hin D Three”) is a type II site-specific deoxyribonuclease restriction enzyme isolated from Haemophilus influenzae that cleaves the DNA palindromic sequence AAGCTT in the presence of the cofactor Mg2+ via hydrolysis. …