A revolution in nucleic acid detection, which can replace the sharp technology of PCR

Biolink report: It has been thirty years since the birth of PCR technology

Recombinase Polymerase Amplification the RPA ( Recombinase Polymerase Amplification ), may alternatively be referred to as a PCR nucleic acid detection techniques

Where is RPA technology sharp

Conventional PCR must go through three steps of denaturation, annealing, and extension, and the PCR instrument is essentially a device that controls the temperature rise and fall

According to reports, RPA detection is highly sensitive and can amplify trace amounts of nucleic acid (especially DNA ) templates to a detectable level, and get about 10 ¹² amplified products

RPA can amplify both DNA and RNA , and it eliminates the need for additional cDNA synthesis steps

Currently, TwistAmp® amplicon in the kit 500bp or less

Basic principles of RPA technology

RPA technology mainly relies on three enzymes: recombinase that can bind single-stranded nucleic acid (oligonucleotide primer), single-stranded DNA binding protein ( SSB ) and strand-displacement DNA polymerase

The protein -DNA complex formed by the combination of recombinase and primer can search for homologous sequences in double-stranded DNA .

Quite flexible detection mode

The basic system of RPA amplification ( TwistAmp® Basic ) contains all the reagents needed for DNA amplification.

By adding different enzymes and probes to this basic system, diversified RPA applications

The RPA base system, patented exo probe technology and exonuclease III combined ( TwistAmp® exo ), data can be read in real time, as fluorescence quantitative PCR as

The RPA basic system, fpg probe technology and ribozyme fpg are combined to form a real-time reporting system ( TwistAmp® fpg )

The combination of RPA basic system and ribozyme nfo ( TwistAmp® nfo ) can support end-point detection of the "sandwich method", such as flow chromatography test strip LFD

In addition, RPA amplification can also be easily multiplexed

Unusual primer design

The key to

When designing RPA primers, the denaturation temperature is no longer a key factor affecting the amplification primers

It should be noted that the current TwistAmp® amplification kits do not provide RNase inhibitors
.
In addition, due to the limitations of current production methods, the RPA reaction is not suitable for the amplification of E.
coli standard laboratory strains
.

Biological Communication Editor: Ye Yu