affinityscript q-pcr cdna synthesis kit  (Agilent technologies)

Journal: Materials

Article Title: 3D-Printing of Hierarchically Designed and Osteoconductive Bone Tissue Engineering Scaffolds

doi: 10.3390/ma13081836

Figure Lengend Snippet: Detection of calcium deposition via alizarin staining on undyed scaffold prototype coated with Saos-2 cells after 21 days: ( a ) left scaffold was seeded in RPMI-medium; ( b ) right scaffold was incubated with osteogenic differentiation medium; and ( c ) PCR results for osteogenic differentiation on Day 21. Saos-2 cells seeded on scaffolds were incubated with RPMI medium and osteogenic differentiation medium. Focus was on cDNA for osteogenic proteins: collagen 1 (Col1), alkalic phosphatase (ALP), and osteocalcin (BGLAP). GAPDH was used as internal standard. The 2 −(ΔΔCt) values are presented.

Article Snippet: Contaminating DNA was removed with a RNase-Free DNase Kit (Qiagen, Hilden, Germany) following the manufacturer´s instructions followed by first strand cDNA synthesis using an AffinityScript PCR cDNA Synthesis Kit (Stratagene, La Jolla, USA).

Techniques: Staining, Incubation

Journal: Frontiers in Pharmacology

Article Title: Searching for Drug Synergy Against Cancer Through Polyamine Metabolism Impairment: Insight Into the Metabolic Effect of Indomethacin on Lung Cancer Cells

doi: 10.3389/fphar.2019.01670

Figure Lengend Snippet: Effect of indomethacin on the expression of SAT1 and levels of spermidine/spermine-N 1 -acetyltransferase (SSAT) in non-small cell lung cancer (NSCLC) cells. A549 and H1299 cells were exposed to indomethacin 0.5 and 1 mM for 24 h. The messenger RNA (mRNA) levels of SAT1 and protein levels of SSAT were evaluated using quantitative PCR (qPCR) and western blotting, respectively. (A) The mRNA levels of SAT1 in A549 cells. (B) Representative blot of the SSAT protein levels in A549 cells exposed to indomethacin. (C) Quantitation of the SSAT levels in A549 cells exposed to indomethacin. (D) The mRNA levels of SAT1 in H1299 cells. (E) Representative blot of the SSAT levels in H1299 cells exposed to indomethacin. (F) Quantitation of the SSAT levels in H1299 cells exposed to indomethacin. * p < 0.05; ** p < 0.01, *** p < 0.0001 and ns, no significant compared with untreated control, calculated using one-way ANOVA and Dunnett’s post-test. Data summarize the results of three independent experiments.

Article Snippet: The RNA obtained was treated with the Turbo DNA-free ® deoxyribonuclease (DNAse) kit (Thermo Fisher Scientific, Vilnius, Lithuania), followed by complementary DNA (cDNA) synthesis using the AffinityScript™ Quantitative PCR (qPCR) cDNA synthesis kit (Agilent Technologies, Cedar Creek, TX, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Quantitation Assay