for real-time quantitative pcr (qrt-pcr) with a capacity of 384 samples in one analysis.
The microsatellite loci were amplified with either Mastercycler Gradient Thermocycler
(Eppendorf North America, Hauppauge, New York) or Arktik Thermocycler
(Thermo Fisher Scientific, Inc.
2003), tul4 gene primer has limit of detection up to 200 fg/ L on BioSeeq thermocycler
platforms but it mainly depend on rise in threshold level, primer ratio, annealing temperature, Taq enzyme and probe level.
(Sensoquest, Gottingen, Germany) was programmed as 94[degrees]C for 5 min, 30 cycles of 94[degrees]C for 25 s, 52[degrees]C for 40 s and 72[degrees]C for 50 s, and a final elongation at 72[degrees]C for 6 min.
Furthermore, we confirmed that the genotyping robustness of the HRMA assay by using 2 commonly used HRMA-capable thermocyclers
, as well as a recently introduced HRMA-capable magnetic induction thermocycler
Reactions were run in a Takara thermocycler
using a step-cycle program.
and then diluted (1:1) with ultrapure water.
It also has to be coupled with a method for product detection--either by opening the reaction tube (risking amplicon contamination of the laboratory) or through application of real-time methods (generally fluorescent in nature, requiring the thermocycler
to incorporate complex optical systems for the illumination and observation of reaction tubes in controlled wavelength bands for detection of weak signals, which require complex computer processing to create meaningful data).
Real-Time PCR thermocycler
conditions was set according to primer annealing temperature and RT-PCR TaqMan kit instructions by Miniopticon Real-Time thermocycler
system as in the following table(3):
have built a solar-powered PCR thermocycler
that pushes the sample analysis onto a phone with an add-on and an app.
Amplification was carried out in gradient thermocycler
(applied biosystem) with initial denaturation 95[degrees]C for 5 min followed by 30 cycles each of denaturation at 95[degrees]C for 1 min, annealing at 65 [degrees]C for 1 min, extension at 72[degrees]C for 1 min with a final extension period of 10 min at 72[degrees]C.
Amplifications were performed with a final reaction volume of 50 [micro]l in a thermocycler
programmed for 3 minutes at 94[degrees]C, followed by 33 cycles of 40 seconds at 94[degrees]C, 45 seconds at 53[degrees]C and 45 seconds at 72[degrees]C with a final extension of 5 minutes at 72[degrees]C.