| 主权项 |
1. A mispriming prevention reagent comprising a nucleic acid molecule comprising, in 5′ to 3′ order:
(i) a first condition-dependent stem region comprising a 5′ terminal covalently linked moiety and a first stem nucleic acid sequence, wherein the first stem nucleic acid sequence is at least 6 nucleotides in length and wherein the 5′ terminal covalently linked moiety comprises a cyclic or polycyclic planar moiety that does not have a bulky portion; (ii) a condition-dependent loop region comprising a loop nucleic acid sequence of at least 3 nucleotides in length; and (iii) a second condition-dependent stem region comprising a second stem nucleic acid sequence and a 3′ terminal covalently linked moiety, wherein the second stem nucleic acid sequence is at least 6 nucleotides in length and is complementary to the first stem nucleic acid sequence and wherein the 3′ terminal covalently linked moiety comprises a cyclic or polycyclic planar moiety that does not have a bulky portion, wherein the 3′ terminal covalently linked moiety is non-identical to the 5′ terminal covalently linked moiety, and wherein the 3′ terminus of the second stem region is non-extendable by a DNA polymerase, wherein the first condition-dependent stem region hybridizes to the second condition-dependent stem region in a temperature dependent manner to acquire a stem-loop hairpin conformation. |
