Assessment of DNAse I Treatment to Rule Out Interference Effects
Sample | + Protein | % Cleaved |
---|---|---|
Control | – | 0% |
S1 | Yes | >99% |
S2 | Yes | >99% |
S1 | No | >99% |
S2 | No | >99% |
QPCR was performed as described (17). Purified DNA extracts (5 ug per sample) were incubated with either 5 (S1) or 10 (S2) units DNAse I following manufacturer recommendations. Samples were incubated with either 5 (S1) or 10 (S2) units DNAse I; those denoted “Yes” were spiked 1:1 with MuLV from source “b,” which contained the highest amount of protein (Table III) in our study, among those preparations studied. No-spike, undigested control samples (control) assessed for shearing effects, of which none were detected. Results demonstrate effective (>99%) cleavage for all DNAse I-treated samples, suggesting negligible interference effects are present when DNAse I is applied under these conditions.