Table 1:

Impact of TCEP in enzyme storage diluent on 3′-labeling activity.

Fig.	Storage Diluent	3′-labeling BoMoC	3′ label	Timing (m)	CV	Mean difference	CI 95%	adjusted p-value	miRNA (n)	3′ precision
3B	pH 6.0, 0.2 mM TCEP	F753A	ddR	90/30	0.87	1.02%	(0.84% - 1.2%)	3.98E-24	902	0.72
3B	pH 6.0, 0.5 mM TCEP	F753A	ddR	90/30	0.91	−0.22%	(−0.42% - −0.03%)	1.00E+00	899	0.71
3B	pH 6.0, 1.0 mM TCEP	F753A	ddR	90/30	0.86	−1.77%	(−1.97% - −1.56%)	4.38E-53	902	0.70
3B	pH 7.5, 0.2 mM TCEP	F753A	ddR	90/30	0.94	1.61%	(1.4% - 1.83%)	1.26E-41	900	0.73
3B	pH 7.5, 0.5 mM TCEP	F753A	ddR	90/30	0.92	1.18%	(0.96% - 1.39%)	3.14E-22	899	0.73
3B	pH 7.5, 1.0 mM TCEP	F753A	ddR	90/30	1.04	0.65%	(0.43% - 0.87%)	5.85E-06	894	0.72
3C	pH 7.5, 0.2 mM TCEP	F753A	ddA	30	0.65	−2.73%	(−3.73% - −1.72%)	1.18E-04	903	0.75
3C	pH 7.5, 0.2 mM TCEP	F753A	ddA	60	0.63	−0.51%	(−1.51% - 0.49%)	1.00E+00	903	0.78
3C	pH 7.5, 0.2 mM TCEP	F753A	ddA	120	0.57	1.98%	(1.01% - 2.94%)	6.08E-02	903	0.80
3C	pH 7.5, 0.2 mM TCEP	W403AF753A	ddA	30	0.64	0.63%	(−0.39% - 1.65%)	1.00E+00	903	0.79
3C	pH 7.5, 0.2 mM TCEP	W403AF753A	ddA	60	0.61	3.37%	(2.36% - 4.38%)	8.18E-08	903	0.81
3C	pH 7.5, 0.2 mM TCEP	W403AF753A	ddA	120	0.59	5.51%	(4.48% - 6.54%)	2.12E-21	903	0.84

Notes: For 3′-labeling time, 90/30 refers to 90 minutes of 3′ labeling with ddATP-followed by a 30-minute ddGTP supplementation. Mean difference in 3′ precision and a 95% confidence interval were computed from a paired, two-sided student’s t-test, Bonferroni adjusted p-values. For Figure 3B, all libraries were compared to a library prepared by F753A diluted in pH 6.0 15 mM DTT diluent buffer and labeled with ddRTP for 90/30 minutes. For Figure 3C, all libraries were compared to a library prepared by F753A diluted in pH 7.5 0.2 mM TCEP diluent buffer and labeled with ddRTP for 90/30 minutes.