A Step One Plus Real-Time PCR System (Applied Biosystems) was used to perform the qPCR experiments using the following cycling protocol: one cycle of 60°C for 2 minutes, one cycle of 95°C for 10 minutes, and 50 cycles of 95°C for 15 seconds and 60°C for 20 seconds.
In the present study, the overall survival and progression-free survival rates in patients receiving two cycles of chemotherapy were better than those in patients receiving one cycle of chemotherapy between all of protocol groups, though we did not show those results.
The reactions were run on a GeneAmp 9700 thermocycler (Applied Biosystems) according to the following protocol; one cycle of 95°C for 15 minutes; 25 cycles of 95°C for 10 seconds, 55°C for 5 seconds, 72°C for 4 minutes.
The reactions were run on a PTC-225 thermal cycler according to the following protocol; one cycle of 95°C for 1 minute; 25 cycles of 95°C for 10 seconds, annealing temperature (64°C for full exon 5 and exon 8, 68°C for exons 6 and 7) for 30 seconds, 72°C for 3 minutes.
according to the following protocol: one cycle of 95°C for 30 s and 25 cycles in the following sequence: 96°C for 10 s, 50°C for 5 s and 60°C for 2 min.
Reactions were run on a Mastercycler gradient thermal cycler (Eppendorf, Lepecq, France) according to the following protocol: one cycle of 96°C for 2 minutes; then 25 cycles of 96°C for 10 seconds, 50°C for 5 seconds and 60°C for 2 minutes.
The cycling protocol started with one cycle of 95°C for 10 minutes for enzyme activation followed by 45 cycles of 95°C for 5 seconds, TA (Table 1) for 10 seconds, 72°C for 10 seconds, one cycle of 95°C for 1 minute, one cycle of 65°C for 1 minute and a melt from 65 to 95°C for all assays.
Response rates were analysed for the intent-to-treat (ITT) population, which was defined as patients who had received at least one cycle of treatment and for the per-protocol population, which was defined as patients assessable for response who had received at least two cycles of chemotherapy.
PCR protocols were one cycle of 95°C for 10 min; 38 cycles of 94°C for 30 s, 50°C (Ots104, Ots107) or 54°C (OMM1128, OMM1231, Ots3 and Omy77) for 30 s, 72°C for 30 s; and a final; extension of 72°C for 10 min. PCR products were run on an ABI 3730 genetic analyzer (Applied Biosystems), and genotypes were visualized with the genotyping software, GeneMapper® 3.7 (Applied Biosystems).
Of the 474 women who were enrolled, 469 received at least one cycle of paclitaxel as specified by the study protocol.
