Quantitative real-time PCR (Q-PCR
) was performed using an ABI Prism 7500 sequence-detection system (Applied Biosystems, Foster City, CA, USA) with SYBR Green Realtime PCR Master Mix (Toyobo, Osaka, Japan), under the manufacturer's instructions.
Table I.- Primer information of candidate gene using q-PCR
To validate gene expression, we randomly chose the upregulated genes HIF-1A, NF-[kappa]B, and BIRC3 and the downregulated gene Bcl2 to do the q-PCR
. The specificity of q-PCR
primers was confirmed by regular PCR (Additional file 3).
experiment was performed in triplicate, with each reaction solution (20 [micro]L) containing 200 nM of each primer, 10 [micro]L of 2X SYBR Green Supermix (BioRad), and 5-30 ng of the DNA template.
. To generate the absolute quantitative standard curve for Q-PCR
analysis on lentivirus copy number measurement, we used the PCR product of the mouse GUS gene and cloned it with the TA cloning vector (pTA[R] Easy Cloning Kit), which was purchased from Genomics BioSci and Tech (Taipei, Taiwan).
(a) Expression of Cadm1 in different developmental stages of mouse inner ear examined by Q-PCR
. The bar graphs show quantification of the PCR results with each value representing the mean [+ or -] standard error.
Expression of IL-10 and TLR4 in T Cell after Transfection by Q-PCR
Moreover, quantitative analysis using q-PCR
revealed that HDF supplemented with 30 [micro]M RA suppressed mRNA expression of iNOS to 20.8% of TNF-[alpha] alone (P<0.05; Figure 3B).
The Ultrasensitive Q-PCR
Assay for EGFR Mutation Detection
Table 1: Gene-specific primers used in Q-PCR
In order to investigate the effect of mechanical injury on ECM metabolism, the ECM production of L929 fibroblast was detected using Western blot and q-PCR
after 5333 [mu]strain.
The authors would also like to thank the Genetics and Molecular Biology Laboratory, Directorate of Poultry Research, Hyderabad, for giving them access to their Q-PCR