CTO | Totalno DNase | Log10 | TotalDNase | Log10 | Difference | |
---|---|---|---|---|---|---|
PRV | a | 2.31 × 109 | 9.4 | 7.51 × 109 | 9.9 | 0.5 |
d | 1.70 × 109 | 9.2 | 6.53 × 108 | 8.8 | 0.4 | |
PRV dilutions | a1:10 | 2.23 × 1010 | 10.4 | 8.86 × 109 | 10.0 | 0.4 |
a1:100 | 3.17 × 1010 | 10.5 | 1.86 × 109 | 9.3 | 1.2 | |
a1:1000 | 3.17 × 1010 | 10.5 | 1.34 × 109 | 9.1 | 1.4 | |
d1:10 | 2.09 × 1010 | 10.3 | n/a | n/a | – | |
d1:100 | 1.62 × 1010 | 10.2 | 6.86 × 108 | 8.8 | 1.4 | |
d1:1000 | 1.73 × 1010 | 10.2 | 5.77 × 108 | 8.8 | 1.4 |
Dilution of virus preparations 1:100 prior to DNAse I digestion ensures the absence of any unwanted enzymatic interference effects. This is most relevant to dsDNA viruses such as PRV, where failure to optimize digestion conditions could in principal yield an overestimation of the true particle-associated genome copy number. PRV preps were either undiluted (top) or diluted 1:10, 1:100, or 1:1000 (bottom) prior to capsid digestion/nucleic acid extraction. Results suggest that a minimal dilution of 1:100 is required to achieve optimal digestion of free nucleic acid.