Table of Contents
What is strand displacement?
Definition: The rejection of the broken 3′ single-strand DNA molecule that formed heteroduplex DNA with its complement in an intact duplex DNA. The Watson-Crick base pairing in the original duplex is restored.
What is BST polymerase?
Bst Polymerase, or Bst DNA Polymerase I, is an enzyme isolated from Bacillus stearothermophilus. This 97-kDa heat-stable enzyme features a similar 5′-3′ polymerase activity of E. coli, but lacks the 3′-5′ exonuclease activity.
What is toehold DNA?
DNA nanotechnology relies on the toehold-mediated strand displacement reaction (SDR). This reaction between two DNA structures initiates at the single-stranded (sticky) end, called a toehold, where the shorter strand in the duplex exchanges with a longer complementary invader strand, requiring no enzymatic mediation.
What is isothermal DNA amplification?
Loop-mediated isothermal amplification (LAMP) is a rugged, low-cost method for specific DNA detection, with a visual readout. LAMP is especially useful in field settings for rapid diagnosis of plant pathogens or infectious disease agents like malaria, Zika, or tuberculosis.
How does a lamp assay work?
In a LAMP assay, the reaction takes place in a single tube containing buffer, target DNA, DNA polymerase and primers. The tube is incubated at 64°C in a regular laboratory water bath or heat block that helps in maintaining a constant temperature.
What is a 3 5 exonuclease?
An associated 3′–5′ exonuclease activity allows a polymerase to remove misincorporated nucleotides, and ensures the high-fidelity DNA synthesis that is required for faithful replication. Proofreading 3′–5′ exonucleases can be divided into intrinsic, polymerase-associated enzymes, or independent autonomous enzymes.
How is Taq polymerase different from DNA polymerase?
Taq polymerase is found in thermophilic bacteria and purified in in vitro DNA replication. The key difference between Taq polymerase and DNA polymerase is that Taq polymerase can withstand high temperatures without denaturing while other DNA polymerases denature at high temperatures (at protein degrading temperatures).
Is Taq a polymerase?
Taq polymerase is the heat-stable (thermostable) DNA polymerase extracted from the thermophilic bacteria Thermus aquaticus. Its predominant function is in the polymerase chain reaction (PCR) technique, where it automates the repetitive step of amplifying specific DNA sequences.
How does strand displacement amplification work?
Strand Displacement Amplification (SDA) is an isothermal, in vitro nucleic acid amplification technique based upon the ability of HincII to nick the unmodified strand of a hemiphosphorothioate form of its recognition site, and the ability of exonuclease deficient klenow (exo- klenow) to extend the 3′-end at the nick …
What is the difference between isothermal amplification and PCR?
Answer. The main distinction between isothermal amplification and PCR is that in the former, amplification is achieved using a constant temperature, while in the latter, cycling of temperatures is required.
What is isothermal PCR used for?
Loop-mediated isothermal amplification (LAMP) LAMP is especially useful in field settings for rapid diagnosis of plant pathogens or infectious disease agents like malaria, Zika, or tuberculosis. Table 2 summarizes the differences between LAMP and PCR. Table 2. Comparison of PCR and LAMP.