The Chemicals, Equipments & Supplies box on the right contains a list of materials used in this protocol. Click on each item for the direct links to order from available suppliers. Quantitative Real Time PCR workflow 4. Designing and validating primers DESIGN: When designing primers, consideration should be given to the area of the target gene being amplified (target sequence) and the thermodynamic properties of the primers themselves. All of these Comparison of the Performance of QF-PCR with QPCR as A Rapid Molecular-Based Method for Sex Chromosome Aneuploidies Detection. by quantitative real-time polymerase chain reaction (PCR) in a Supplementary Table S1. Sequences for RT-PCR and RT-qPCR primers. RT-PCR Primers Product Size (bp) Forward Sequence Reverse Sequence TLR4 206 5'-GACAACCAGCCTAAAGTATT-3' 5'-TGCCATTGAAAGCAACTCTG-3' Test 'Touchdown cycling' as an alternative for multiplex PCR reactions. Touchdown cycling is a common feature on many thermocyclers (consult your manual for instructions). This feature alters the temperature incrementally during the annealing step to accommodate primer pairs with different optimal annealing temperatures. Previous article in issue: Microbial diversity of mid-stage Palinurid phyllosoma from Great Barrier Reef waters A TaqMan real-time polymerase chain reaction (PCR) was used to detect DNA of piroplasms in specimens of clotted blood or EDTA blood samples of the same animals. Out of the 105 serum samples, the IFAT detected antibodies against T. equi in 35 (33.3%) cases while the cELISA gave 34 (32.4%) positive results. Clinical research suggests a role for viral load measurement in predicting and monitoring Epstein-Barr virus (EBV)-associated diseases. The aim of this study was to assess the performance of the recently commercially available quantitative assays for EBV based on real-time PCR: the RealArt EBV LC PCR kit and the LightCycler EBV quantification kit. monocytogenes in other real time PCR based stu