Skip to main content
. Author manuscript; available in PMC: 2022 Nov 1.
Published in final edited form as: J Immunol Methods. 2021 Aug 28;498:113132. doi: 10.1016/j.jim.2021.113132

Table 1A.

RNA purity, yield, and absolute copy number of 18S rRNA in urine samples processed using the CCBP or the ZFBP.

S.no Volume (cc)a A260/280 Ratiob total RNA (μg)d 18S rRNA copies/μg RNAf
CCBP ZFBP CCBP ZFBP ΔA260/280 Ratio c CCBP ZFBP Δtotal RNA e CCBP ZFBP
1 30 30 1.94 2.03 0.09 2.33 4.51 2.18 1.84E+10 1.19E+10
2 50 25 2.05 2.04 n/a 1.31 2.41 n/a 1.83E+10 6.40E+09
3 50 50 1.19 1.96 0.77 0.13 1.31 1.18 1.98E+08 1.75E+09
4 50 30 2.15 2.08 n/a 1.36 2.46 n/a 2.54E+10 8.97E+09
5 35 20 2.11 1.97 n/a 0.70 2.70 n/a 7.94E+09 3.96E+09
6 70 30 2.08 1.96 n/a 2.55 12.83 n/a 2.57E+10 1.86E+09
7 50 30 1.98 1.97 n/a 1.32 2.43 n/a 2.32E+10 3.49E+09
8 50 50 3.94 1.19 2.76 0.10 0.18 0.08 1.58E+08 2.47E+08
9 50 20 2.13 2.13 n/a 7.68 2.43 n/a 9.55E+09 9.02E+09
10 40 40 2.11 1.84 0.27 0.30 0.47 0.17 1.52E+09 3.70E+09
11 70 30 1.87 2.06 n/a 0.51 1.12 n/a 1.06E+10 1.50E+09
12 50 50 1.37 1.98 0.61 0.09 0.35 0.26 4.87E+08 8.63E+07
13 50 50 1.96 2.07 0.11 1.95 2.58 0.63 1.77E+10 2.60E+09
14 70 40 1.99 2.00 n/a 1.54 3.50 n/a 2.79E+10 2.82E+09
15 50 50 1.96 1.86 0.10 0.61 0.86 0.25 3.54E+09 4.18E+08
16 70 40 2.10 2.00 n/a 4.02 3.27 n/a 7.82E+10 7.89E+08
17 50 50 2.04 2.47 0.44 0.68 0.33 0.35 7.09E+09 4.47E+08
18 50 50 1.62 2.51 0.89 0.32 0.36 0.04 1.52E+09 3.58E+08
19 45 45 1.87 1.87 0.00 0.45 0.31 0.14 2.92E+09 2.31E+08
20 40 40 2.61 2.01 0.61 0.14 0.74 0.60 3.34E+08 6.58E+08
21 43 43 2.07 2.18 0.11 0.11 0.39 0.28 3.40E+09 2.59E+08
22 50 50 1.86 2.04 0.18 0.04 7.90 7.86 8.65E+07 8.32E+09
23 45 45 1.84 1.99 0.16 0.30 3.94 3.64 2.29E+10 2.00E+10
24 30 30 1.94 2.06 0.12 0.80 2.65 1.85 5.49E+09 1.08E+10
25 37 37 1.49 2.07 0.58 0.13 0.19 0.06 9.69E+07 1.54E+08
26 40 40 1.63 1.79 0.16 0.12 0.24 0.12 1.44E+08 1.15E+08
27 50 50 1.21 1.94 0.73 0.13 0.37 0.24 1.45E+08 1.99E+08
28 45 45 2.04 1.89 0.15 0.15 1.39 1.24 1.28E+08 3.02E+09
29 40 40 2.16 2.23 0.07 0.16 0.18 0.02 8.47E+08 3.10E+08
30 50 50 1.57 1.73 0.16 0.07 0.40 0.33 1.86E+08 2.93E+09
31 50 50 1.38 1.85 0.47 0.11 0.20 0.09 1.49E+08 9.04E+07
32 47 47 1.53 1.96 0.43 0.10 0.59 0.49 6.49E+07 1.86E+08
33 30 30 1.32 2.00 0.69 0.20 1.31 1.11 2.13E+08 2.94E+08
34 45 45 1.00 1.99 0.99 0.05 0.24 0.19 7.21E+07 2.41E+08
35 50 50 2.26 1.97 0.29 0.62 1.93 1.31 3.07E+10 8.80E+08
36 30 30 2.09 2.06 0.03 0.07 0.81 0.74 1.51E+08 2.34E+09
37 45 45 1.56 2.06 0.50 0.09 0.98 0.89 9.50E+07 9.69E+09
38 35 35 1.61 2.00 0.39 0.10 0.59 0.49 8.35E+07 1.75E+08
39 50 50 1.70 1.90 0.21 0.34 0.39 0.05 2.12E+09 3.49E+08
40 40 40 1.51 2.10 0.59 0.09 0.31 0.22 1.34E+08 1.06E+09
41 50 50 1.54 1.65 0.11 0.09 0.25 0.16 1.45E+08 1.14E+09
42 45 45 1.26 1.30 0.03 0.10 0.26 0.16 4.96E+08 1.53E+08
43 30 30 1.98 2.04 0.06 0.15 0.46 0.31 1.46E+08 2.67E+08
44 30 30 1.98 2.07 0.09 0.54 2.10 1.56 1.03E+10 1.17E+10
45 35 35 1.99 2.08 0.10 0.42 1.52 1.10 2.06E+09 1.70E+09
46 25 25 2.12 2.11 0.01 0.84 0.96 0.12 1.40E+10 1.40E+10
47 30 30 2.02 2.52 0.51 0.43 0.17 0.26 1.82E+09 7.37E+07
N=47g Median 45 40 1.96 2.00 n/a 0.30 0.81 n/a 1.52E+09 1.06E+09
25th Percentile 37 30 1.56 1.94 n/a 0.10 0.33 n/a 1.46E+08 2.47E+08
75th Percentile 50 50 2.08 2.07 n/a 0.70 2.43 n/a 1.06E+10 3.70E+09
Mann-Whitney P value 0.03 0.06 <0.0001 0.80
N=38h Median 45 1.87 2.00 0.20 0.15 0.47 0.30 4.11E+08 4.33E+08
25th Percentile 35 1.53 1.89 0.10 0.10 0.30 0.16 1.45E+08 2.23E+08
75th Percentile 50 2.04 2.07 0.58 0.44 1.33 1.10 3.44E+09 2.95E+09
Wilcoxon P value 0.0007 < 0.0001 0.79
a

The volume of paired urine samples processed under each protocol. CCBP: Total RNA was isolated using RNeasy Mini Kit (Qiagen, Cat. 74104). ZFBP: Total RNA was isolated using Urine RNA Isolation Kit (Zymo, Cat. R1038/R1039 Ver. 1.0.0).

b, d

The quantity (absorbance at 260nm) and purity (ratio of absorbance at 260 and 280nm) of RNA isolated from the urine cell pellet (CCBP) or filtrate (ZFBP) were measured using the Nanodrop® ND-1000 UV-spectrophotometer.

c,e

The absolute difference between the CCBP and ZFBP of the A260/280 ratio and of the total RNA yield.

f

Absolute copy number of 18S rRNA was measured using gene specific oligonucleotide pairs and TaqMan probe in customized RT-qPCR assays on ABI 7700 Detection System (Applied Biosystems).

g

All 47 paired urine samples included in data analyses; P values derived using Mann-Whitney test.

h

38 of 47 paired urine samples with equal volumes included in the analysis. The shaded rows (urine sample pairs 2, 4, 5, 6, 7, 9, 11, 14 and 16) were not split evenly and were excluded from data analyses; P values derived using Wilcoxon matched pairs signed-ranks test.