What is isothermal DNA amplification?

In contrast to PCR, isothermal amplification enables rapid and specific amplification of DNA at constant temperature (60-65 °C) avoiding the requirement of thermal cycling. Phi29 DNA polymerase is the enzyme of choice for whole genome amplification (WGA). …

What is isothermal amplification methods?

Isothermal amplification methods provide detection of a nucleic acid target sequence in a streamlined, exponential manner, and are not limited by the constraint of thermal cycling. Although these methods can vary considerably, they all share some features in common.

How are isothermal amplification technologies different from PCR technology?

In contrast to the polymerase chain reaction (PCR) technology, in which the reaction is carried out with a series of alternating temperature steps or cycles, isothermal amplification is carried out at a constant temperature, and does not require a thermal cycler.

What is LAMP technique?

“LAMP” which stands for Loop-mediated Isothermal Amplification is a simple, rapid, specific and cost-effective nucleic acid amplification method solely developed by Eiken Chemical Co., Ltd.

How does multiple displacement amplification work?

The amplification reaction initiates when multiple primer hexamers anneal to the template. When DNA synthesis proceeds to the next starting site, the polymerase displaces the newly produced DNA strand and continues its strand elongation.

Is real time PCR quantitative?

Real-time PCR results can either be qualitative (the presence or absence of a sequence) or quantitative (copy number). Quantitative real-time PCR is thus also known as qPCR analysis. In contrast, PCR is at best semiquantitative.

Which is better Elisa or PCR?

Compared to ELISA, real-time PCR showed greater agreement among duplicate samples. ELISA was found to be less time consuming and easier to perform than real-time PCR. ELISA and real-time PCR showed 100% specificity during reference sample testing.

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 …

How is Recombinase polymerase used in isothermal amplification?

Recombinase polymerase amplification is a low temperature DNA and RNA amplification technique. Instead of melting DNA strands apart at high temperatures, isothermal amplification takes advantage of DNA polymerases with high strand displacement activity, like Bst or Phi29 DNA polymerases.

How is loop-mediated isothermal amplification ( LAMP ) used for DNA detection?

Loop-mediated isothermal amplification (LAMP) 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 are primers used in isothermal amplification systems?

Isothermal amplification systems can use sequence-specific primers to detect target genes, or random primers for whole genome amplification. Loop-mediated isothermal amplification (LAMP) Loop-mediated isothermal amplification (LAMP) is a rugged, low-cost method for specific DNA detection, with a visual readout.

How long are the products of isothermal amplification?

Products of the reaction are extremely long (>30 kb) and highly branched through the multiple displacement mechanism.