Primer Premier

[…] ace qpcr rt kit (toyobo, japan) for cdna synthesis. the cucumber genome database was applied for searching the full-length coding sequences (cds) of csdir16 gene (gene id csa4m280630.1), and primer premier 5.0 (premier biosoft international, ca, usa) was used to design the specific primers for cloning the full-length cds. the full-length csdir16 orf was amplified by pcr using the primers […]

[…] 7500 real-time pcr system (applied biosystems, usa) using kapa sybr fast universal qpcr kit (kapa biosystems, usa). specific primers of the genes shown in supplementary table  were designed using primer premier 5 and confirmed by oligo 6.0. using β-actin as a reference, relative expression levels of the genes were quantified using 2(−δδct) methods., go and kegg pathway enrichment analysis […]

[…] the significantly different sequences between the nun moth, casuarina moth, and asian gypsy moth in the shared fragment, aiming to design the specific primers., according to the sequencing results, primer premier 6.0 software (http://www.premierbiosoft.com/primerdesign/) was used to design and screen‐specific pcr primer pairs with a pcr product size of 150–300 bp, the primer length ranges […]

[…] topological overlap was calculated to measure network interconnectedness., two candidate genes screened from the wgcna modules were further analyzed by sanger sequencing. primers were designed using primer premier 6 (http://www.premierbiosoft.com/) software, and homologous genes were cloned from k14, kt1 and tri. protein sequences of the cloned genes were compared with their homologs […]

[…] was performed using a prime-script™ ii first strand cdna synthesis kit (takara bio, dalian, china) according to the manufacturer’s instructions. the primer sets for each unigene were designed by primer premier 5.0, and their sequences are listed in . qrt-pcr was carried out on an abi 7500 real-time pcr system (thermo fisher scientific, inc., waltham, ma, usa) with sybr premix ex taq™ ii kit […]

[…] 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 […]

[…] 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 […]

[…] 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 […]

[…] functional substances in e. ferox seed were selected to validate the transcriptome data using quantitative real-time pcr. the gene-specific primers for the six unigene sequences were designed with primer premier 5 software [], and the β-actin gene was used as an internal gene expression control: the gene was amplified with forward primer 5′-gactctggtgatggtgt-3′ and reverse primer […]

[…] and corresponding inbred lines using the trizol reagent (invitrogen, usa). primers specific for several genes related to the phenylpropanoid and benzoxazinoid metabolic pathways were designed using primer premier 5.0 for a quantitative real-time polymerase chain reaction analysis (qrt-pcr) (table ). primers were also designed for the actin gene, which was used as an internal standard […]