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The first PCR in the 5' RACE (3' RACE) was performed using the 5' RACE (3' RACE) outer primer supplied in the kit (Ambion) and primers specific for the respective subunit (Table S2).
We used 5' RACE Outer and Inner forward primers corresponding to the 5' RACE adapter sequence (Ambion), in combination with an ALMS1-specific reverse primer spanning the exon 4/5 junction (5'-aggaattcccctcagaggtgcaaaacttag-3') and a nested reverse primer in exon 4 (5'-caaattcttggtcttgtgtcaaac-3') respectively.
Mathematical expressions were derived for inner race, outer race and rolling element local defects.
PCR was carried out as described in 5′ RLM-RACE using only outer primer (3′ RACE Outer Primer).
Waviness of all the contact surfaces (including inner race, outer race, and rollers) is included and compared in this analysis.
The elastic deformations at the interfaces between gear and shaft, inner race and outer race, outer race and housing are formulated by the corresponding contact stiffness.
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The Outer 3′-RACE PCR was first performed with primer P2-S (Table 1) and 3′-RACE Outer Primer (provided in the kit) under the conditions of 94 °C for 3 min followed by 35 cycles of amplification (94 °C for 30 s, 56 °C for 30 s, and 72 °C for 2 min), and a final elongation at 72 °C for 10 min.
The RNA was then reverse transcribed using oligo-d(T) as above, and cDNA was amplified with an antisense gene-specific primer (Table 1: feCath 5'-RACE), based on the feCath sequence, and a 5'-RACE Outer Primer (Table 1: 5'-RACE Outer Primer).
Single point faults were introduced separately at the inner-race, outer-race, and rolling element (i.e., ball) of the test bearing using electro-discharge machining with fault diameters of 7 mm.
Four different categories include; healthy, inner race defect, outer race defect, and double holes in outer race are investigated.
The maximum attenuation occurs at the transmission from inner race to outer race, while the minimum is between the outer race and the housing.
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