Table 2. Filter binding analysis of Kd with HIV RT wild type and RNase H minus.
| 1Duplex sequence | Name | 2Kd wt (nM) | 2Kd E>Q (nM) |
| 5′-AAAAGAAAAGGGGG-3′ (r or d) 3′-TTTTCTTTTCCCCCTGAC-5′ (d) | HIV DNA/RNA PPT | DNA- 132±30 RNA- 1290±70 | DNA- 44±12 RNA- 180±90 |
| 5′- GAGCGCGCGGCC-3′ (r) 3′-GGACTCGCGCGCCGGCGAC-5′ (d) | Ty3 RNA PPT | 2380±840 | 850±425 |
| 5′-CCUGAGCGCGCGGCC-3′ (r) 3′-GGACTCGCGCGCCGGCGAC-5′ (d) | Ty3+3 RNA PPT | 900±205 | 105±49 |
| 5′-AAAAGAAAAGCGCGC-3′ (r) 3′-TTTTCTTTTCGCGCGTGAC-5′ (d) | Control-1 | 1760±290 | 120±37 |
| 5′-AAUUCGAGCUCGGTA-3′ (r) 3′-TTAAGCTCGAGCCATTGAC-5′ (d) | Control-2 | 2610±710 | 1090±170 |
1
Duplexes were prepared by hybridization and gel purification of the hybridized material as described in the Methods section. r, RNA strand; d, DNA strand.
2
Results are an average of 2–4 independent experiments ± standard deviation. wt, wild type RT; E>Q, RNase H minus E478>Q RT. Assays with wt were without Mg2+ while 6 mM Mg2+ was present with E>Q. Values were calculated based on protein mass as provided by the manufacturer (wt) or determined as described (E>Q) [34].