Table 3A.
Performance characteristics of RNA isolated from the filtrates stored at −80°C or shipped to the GEM Core at Ambient (Room) Temperature.
| S.no | Volume (cc)a | A260/280 Ratiob | Nanodrop based total RNA (μg)c | Qubit based total RNA (μg)d | 18S rRNA copies/ μg RNAe | TGFpl copies/ μg RNAe | has-miR-26a copies/ μg RNAe | ||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 80°C | RT | 80°C | RT | 80°C | RT | −80°C | RT | −80°C | RT | −80°C | RT | ||
| 1 | 50 | 1.85 | 2.00 | 2.94 | 2.57 | 1.01 | 1.07 | 2.36E+08 | 2.70E+08 | 1.42E+03 | 2.87E+03 | 2.82E+06 | 2.36E+06 |
| 2 | 50 | 2.06 | 2.13 | 0.83 | 0.79 | 0.42 | 0.44 | 1.32E+09 | 8.86E+08 | 9.30E+03 | 3.64E+03 | 9.57E+05 | 1.16E+06 |
| 3 | 50 | 2.04 | 2.01 | 1.83 | 1.59 | 1.39 | 1.32 | 1.92E+07 | 3.02E+07 | 1.68E+03 | 6.62E+02 | 1.00E+07 | 1.08E+07 |
| Median | 50 | 2.04 | 2.01 | 1.83 | 1.59 | 1.01 | 1.07 | 2.36E+08 | 2.70E+08 | 1.68E+03 | 2.87E+03 | 2.82E+06 | 2.36E+06 |
| 25th Percentile | 50 | 1.85 | 2.00 | 0.83 | 0.79 | 0.42 | 0.44 | 1.92E+07 | 3.02E+07 | 1.42E+03 | 6.62E+02 | 9.57E+05 | 1.16E+06 |
| 75th Percentile | 50 | 2.06 | 2.13 | 2.94 | 2.57 | 1.39 | 1.32 | 1.32E+09 | 8.86E+08 | 9.30E+03 | 3.64E+03 | 1.00E+07 | 1.08E+07 |
| Wilcoxon P value f | 0.50 | 0.25 | >0.9999 | >0.9999 | 0.75 | 0.75 | |||||||
Urine samples were processed using the ZR Urine RNA Isolation Kit™ and the filtrates were stored either at −80°C or shipped overnight to the GEM Core. Total RNA was isolated from the filtrates and analyzed.
The volume of urine samples processed.
A260/280 ratio measured using the NanoDrop® ND-1000 spectrophotometer.
The amount of RNA measured using the NanoDrop® ND-1000 spectrophotometer.
The amount of total RNA measured using the Qubit fluorometer.
Absolute copy numbers of 18S rRNA, TGF-β1 mRNA, and hsa-miR-26a miRNA were measured using customized RT-qPCR assays using gene specific oligonucleotide pairs and TaqMan probes on 7500 Fast Real-Time PCR System.
P values derived using Wilcoxon matched pairs signed-ranks test.