The arrays were then stained with streptavidin-phycoerythrin using the standard antibody amplification protocol (GeneChip® Expression Manual, Affymetrix, Inc., Santa Clara, CA).
We next determined whether zTERT-MO1 inhibits telomerase activity in vivo using the standard telomere repeat amplification protocol (TRAP) assay.
FISH positivity was estimated using the standard Colorado criteria (gene amplification; 2≦ MET gene (red)/ CEP7q (green) per cell plus high polysomy; 5 copies≦mean MET per cell) [ 9, 11- 13].
The researchers checked that the DNA released this way not only retained the same information – the same sequence – but could also be copied using the standard laboratory method for DNA amplification, called the polymerase chain reaction (PCR).
Inter-assay variability was calculated using the standard deviation of the six amplification values (Cq and copy values).
Spoligotyping of 38 samples from study 2 was performed at VLA using the standard protocol (35 cycles of amplification).
Using the standard curve method generated by amplification of 0.08 50 ng of LNCaP cDNA, the resulting Ct values were converted to picogram quantities.
Therefore, we recommend using the standard protocol instead of using the amplification strategy for samples down to 1 μg of total RNA, and our simplified amplification protocol for smaller samples, at least down to 0.1 μg.
Differences in amplification were determined using the standard curve method.
Analysis of EGFR amplification was performed using the standard dual-colour EGFR Spectrum Orange™/CEP7® Spectrum Green™ probe (Visys, Downers Grove, IL USA).
