Primer Premier

[…] TaKaRa, Dalian, China). The T. elegans actin gene (Genbank accession no. MG322127) was used as control. Each reaction was performed with 200 ng/μl cDNA sample. Primers for RT-qPCR were designed using Primer Premier 5.0 software and are listed in Table . The RT-qPCR reactions were conducted in 20 μL reaction mixtures containing 10 μL SYBR Premix Ex Taq II, 20 ng cDNA templates, 0.2 μM of each prime […]

[…] alignment of these sequences was performed with Clustal X (version 2.0) and the conserved region was selected as the target for the primer design (Fig. ). The primer design was then carried out using Primer Premier (version 6.0, Premier Biosoft International, USA). The McrA 997 F primer was newly designed in this study and the efficiency was confirmed in silico with the NCBI Primer-Blast tool.Figu […]

[…] r the qRT-PCR experiment. And 12 genes with different expression levels were randomly selected for validating the expression results of RNA-seq sequencing. Primers (Additional file ) were designed by Primer Premier 5(http://downloads.fyxm.net/download-now-Primer-Premier-Others-Home-&-Education-101178.html) based on the selected unigene sequences. PCR reaction mixture was consisted of 7 μl ddH2O, 1 […]

[…] 2 × SYBR Master Premix ExTaq (Takara), 1 μl cDNA template (1:5 dilution), and 1 ul of each forward and reverse primer for the selected gene of interest. All primers were designed based on cDNAs using Primer Premier 5.0 (Additional file : Table S1). Three biological and three technical relipcates per sample were analysed using Bio-Rad Real-Time System (BioRad, Hercules, CA, USA) based on the 2-ΔΔCT […]

[…] r 10 s and annealing at 60 °C or 55 °C (Additional file ) for 30 s. All amplifications were followed by dissociation curve analysis of the amplified products. Specific primers were designed using the Primer Premier (v5.0), and specificities were confirmed with BLAST. The gene expression levels were normalized with the reference gene GAPDH by using 2−ΔΔCt value methods. The data were presented as t […]

[…] ned using a NanoDrop 2000C spectrophotometer (Thermo Scientific, USA). Reverse transcription was conducted using PrimeScript™ RT reagent kit (TaKaRa, Japan). Gene specific primers were designed using Primer Premier 6 (PREMIER Biosoft Int, USA) based on SmDML coding sequences. SmUBQ10 was used as an internal control as described previously. The expression of Smi-miR7972a and Smi-miR7972b was analyz […]