G02, H02, G03, H03, G04, H04 were the results of when primers concentration were 0.75 μmol/L and probe concentration was 0.5, 0.75, 1 μmol/L, respectively A01 B01 C01 D01 E01 F01 G01 H01 G02 H02 G03 H03 G04 H04 0 20000 40000 60000 80000 100000 120000 140000 160000 Event ...
Description ddPCRSupermix orProbes(NodUTP)isa2xconcentrated, ready-to-usereactioncocktailcontainingallcomponents— exceptprimers,probe(s),andtemplate—required orprobe- basedDropletDigital ™ PCR(ddPCR).Themixturedelivers maximumtargetspecicityandfuorescenceamplitude andminimumdropletvariabilitytoensureprecisetarg...
Component Volume per Reaction, µl Final Concentration 2x ddPCR Supermix for Probes (No dUTP) 20x target primers/probe (FAM or Cy5)* 20x target primers/probe (HEX/VIC or Cy5.5)* Diluted restriction enzyme (see DNA Digestion section) Sample RNase-/DNase-free water Total volume*** 10 1...
The kit contains ddPCR Supermix for Probes, primers, wild-type probes, mutant probes, positive controls and negative controls. It combines the superior product of Kebai Bio-molecular diagnostic reference materials with ddPCR probe primers to provide customers with Provide a more comprehensive and co...
Joke Verreth, Bram De Wilde, Tom Van Maerken, Katleen De Preter,et al.mix concentration). Each 20 µl dPCR reaction contains 10 µl 2 ×ddPCR Supermix for Probes(Bio-Rad,1863010), 2 µl of primer and probe mix (with a total of 4 primers and 2 probes per duplexassay) and...
Finally, in the multiplex modules, variations in primers and probe concentration were performed to differentiate clusters of positive droplets in a single detector channel. The best differences were achieved at concentrations with 1:3 differences. The results showed...
those used for most standard TaqMan probe-based assays. Following PCR, each droplet is analyzed or read to determine the fraction of PCR-positive droplets in the original sample. These data are then analyzed usingPoisson statisticsto determine the target DNA template concentration in the original ...
A T7 promoter sequence was added to the 5′ primer sequence. As a result, a 3823 bp DNA fragment (Supplemental Fig. 1A) was amplified and purified with Zymo DNA clean and a concentration kit (Cat# D4033, Zymo Research, irvine, CA) (Supplemental Fig. 1B). Utilizing the T7 promoter ...
Then, the relative concentration of targets, primers and probes are higher, while the inhibitor is relatively lower, which perfectly overcoming the shortcomings of conventional qPCR [23]. Our study had several limitations. First, due to the specialized instruments and consumables, ddPCR has a ...
Here, we tested and compared the performance of two primer/probe assays to detect and quantify the abundance of bullfrogs in Western Europe in silico and in situ using digital droplet PCR (ddPCR). Although both assays proved to be equally target-specific and sensitive, one outperformed the ...