Table 2.
oligonucleotide a |
Tm (Tm)b |
|||
---|---|---|---|---|
IQc | AFc | AAFd | ||
a | 5 ′ -GGC AGG TGG TG-3 ′ (1) | 51° (−9°)e | ||
3 ′ -CCG TCC ACC AC-5 ′ | ||||
b | 5 ′ -CTC GGC GCC ATC-3 ′ (2) | 58° (−7°)f | 57° (−8°)f | |
3 ′ -GAG CCG CGG TAG-5 ′ | ||||
c | 5 ′ -CTC GGC GCC ATC-3 ′ (3) | 60° (−5°)f | 56° (−9°)f | |
3 ′ -GAG CCG CGG TAG-5 ′ | ||||
d | 5 ′ -CTC GGC GCC ATC-3 ′ (4) | 61°(−4°)e | 52°(−13°)f | |
3 ′ -GAG CCG CGG TAG-5 ′ | ||||
e | 5 ′ -ACC GGC GCC ACA-3 ′ | 51° (−10°)g | ||
3 ′ -TGG CCG CGG TGT-5 ′ | ||||
f | 5 ′ -ACC GGC GCC ACA-3 ′ | 48° (−13°)g | ||
3 ′ -TGG CCG CGG TGT-5 ′ | ||||
g | 5 ′ -ACC GGC GCC ACA-3 ′ | 48° (−13°)g | ||
3 ′ -TGG CCG CGG TGT-5 ′ | ||||
h | 5 ′ -CTC GGC GCC ATC-3 ′ (4) | 48°(+10°)e | 44° (+6°)f | |
3 ′ -GAG CC- -GG TAG-5 ′ | ||||
i | 5 ′ -ACC GGC GCC ACA-3 ′ (4) | 49° (+15°)g | ||
3 ′ -TGG CC- -GG TGT-5 ′ | ||||
j | 5 ′ -CTC GGC GCC ATC-3 ′ (4) | 55° (+4°)f | ||
3 ′ -GAG CCG -GG TAG-5 ′ |
G is the C8-modified dGuo.
ΔTm= Tm (modified) — Tm (unmodified). The Tm values for the unmodified Ras-12 (entry a, 11-mer), our NarI (entries b—d, 12-mer), and Fuchs’ NarI (entries e—g, 12-mer) oligonucleotides were 60, 65, and 61 °C, respectively. The Tm values for our unmodified NarI (entry h) and Fuch’s unmodified NarI (entry i) oligonucleotide opposite a two-base deletion were 38 and 34 °C, respectively. The Tm values for our unmodified NarI (entry j) oligonucleotide opposite a one-base deletion was 51 °C.
Conditions: 10 mM phosphate buffer (pH 7.0) containing 100 mM NaCl, 0.05 mM EDTA, and 0.5 A260/mL of each oligonucleotide. The temperature was raised 1 °C min−1.
Conditions: 10 mM Tris, 50 mM NaCl, and 1 mM EDTA; DNA concentration was 40 µg/mL.
Ref 33.
This work.
Ref 40.