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The assay started with 5 µg of total RNA sample.
Day 1 of the lifespan assay started 68 h after seeding the animals into plates.
As shown in Fig. 1B, cell viability (MTT assay) started to decrease at 48 h after DHA treatment, and approximately 80% of the cells lost viability at 72 h.
Briefly, the assay started from 2 to 5 µg total RNA sample, which was size fractionated using a YM-100 Microcon centrifugal filter (from Millipore) and the small RNAs (<300 nt) isolated were 3'-extended with a poly(A) tail using poly(A) polymerase.
The assay started with 5 µg total RNA pools containing equal amount of RNA from individual lung (n = 6 per pool), which was size-fractionated using a YM-100 Microcon centrifugal filter (Millipore, Billerica, MA), and the isolated small RNAs (<300 nt) were 3'-extended with a poly(A) tail using poly(A) polymerase.
Briefly, the assay started with 8 μg of total RNA.
Similar(46)
The overall cost calculated for one PCR assay starting from DNA isolation in this study was minimal, only Rs 230 (less than $5) which is quite similar to cost of one rk39 strip.
Another sensitivity assay starting from quantitatively diluted culture supernatants was also performed (Table 4).
The precision of the Prosigna assay starting from RNA was assessed with 5 pooled breast tumor RNA samples each tested 36 times at each of the three sites.
These SN titres rose slowly until they could be detected with the classical SN assay starting from 4 weeks after inoculation (Pogranichnyy et al., 2000) [ 24].
PCR amplification was performed with Universal Probe Library fluorescent probes (Roche, Basel, Switzerland) and a 5′-nuclease assay, starting with 200 ng of total RNA.
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