Direct PCR (dPCR) is a method of DNA amplification directly from an animal or plant tissue sample without performing DNA isolation and purification steps. This technique greatly reduces experimental time, and cost in genotyping and high-volume projects. It also provides a better option when faced with the challenges of amplifying a very small quantity of sample where the purification step could potentially lead to sample loss.
In many ways, dPCR works like conventional PCR. Both techniques depend on template DNA, primers and a master mix or PCR reagents, and both use a thermal cycler for amplification. The primary difference between dPCR and conventional PCR is the tailored buffers used in dPCR. The modified elements of dPCR enable robust amplification despite the presence of PCR inhibitors often found in crude samples.
ALS SARS-CoV-2 RT-PCR 4G has been developed and produced entirely at ALS in Portugal and is designed mainly for export. This new kit is able to detect all known SARS-CoV-2 variants, including British, Brazilian and South African variants, without cross-reacting with other known related viruses or common respiratory pathogens. The choice of the genomic regions used as targets and the sequences of the primers and hydrolysis probes were based on WHO, CDC and DGS recommendations.
The ALS SARS-CoV-2 RT-PCR 4G kit is an in vitro diagnostic medical device (CE-IVD) that uses real-time PCR nucleic acid amplification technology for the detection of SARS-CoV-2 in clinical samples from the upper respiratory tract of individuals who may or may not be suspected of having COVID-19.
The kit uses the RT-PCR technique, combining amplification of certain regions of the viral genome with its detection by specific hydrolysis probes. The SARS-CoV-2 genome target regions are highly conserved and correspond to two specific genes: ORF polyprotein (ORF1ab gene) and nucleocapsid phosphoprotein (N gene). The kit can also detect the gene that codifies the structural protein of the envelope (gene E) specific to the Sarbecovirus subgenus.
Direct PCR Reagent
Advantages
Rapid amplification: The amplification rate of the polymerase is 6kb/min. 1kb fragment can be amplified within 25min.
Long fragment amplification: For plasmid, λ DNA and other easy templates, the polymerase can effectively amplify > 20kb. For genome, the polymerase can effectively amplify > 8kb. And for cDNA, the polymerase can effectively amplify > 8kb.
High specificity: With hot start technology, the polymerase is 100% inactive below 50°C, and can only be restored by heating at 95°C for 5min.
High tolerance to impurities: Samples of whole blood, serum, cultured cells and urine can be directly amplified without prior DNA purification.
Unit Definition
One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmol dNTP into an acid insoluble substance within 30 min at 74°C.
Description: The Mouse Direct PCR Kit provides a fast preparation and PCR amplIFication that is specIFically designed for mouse genotyping. Buffer L and Protease Plus rapidly digests mouse tissue to release intact genomic DNA that can be used directly as the template for PCR amplIFication.
Description: The Mouse Direct PCR Kit provides a fast preparation and PCR amplIFication that is specIFically designed for mouse genotyping. Buffer L and Protease Plus rapidly digests mouse tissue to release intact genomic DNA that can be used directly as the template for PCR amplIFication.
Description: The Mouse Direct PCR Kit provides a fast preparation and PCR amplIFication that is specIFically designed for mouse genotyping. Buffer L and Protease Plus rapidly digests mouse tissue to release intact genomic DNA that can be used directly as the template for PCR amplIFication.
Description: The Mouse Direct PCR Kit provides a fast preparation and PCR amplIFication that is specIFically designed for mouse genotyping. Buffer L and Protease Plus rapidly digests mouse tissue to release intact genomic DNA that can be used directly as the template for PCR amplIFication.
