Primers for nucleic acid extension or amplification reactions
Disclosed are methods and compositions for use in nucleic acid amplification or extension reactions.
1. A method of assaying for a nucleic acid sequence, the method comprising contacting a split primer with a sample that may comprise DNA derived from a Dengue virus under conditions necessary for the amplification, wherein the split primer has two regions comprising a first sequence complementary to a first region of a target nucleic acid sequence and a second sequence complementary to a second region of a target nucleic acid, and wherein the two regions of the split primer are not separated by an intervening base, and wherein the split primer targets two or more unique locations on a target nucleic acid from the Dengue virus, and wherein the two or more unique locations are separated by at least one nucleic acid base, and wherein amplification of the target sequence is carried out in the presence of a polymerase and a reverse primer under conditions necessary for amplification or extension, and is indicative of the presence of DNA derived from Dengue virus in the sample.
2. The method of claim 1 , wherein the two or more unique locations are separated by, at least three nucleic acid bases.
3. The method of claim 1 , wherein the two or more unique locations are separated by at least five nucleic acid bases.
4. The method of claim 1 , further comprising detecting a product from the amplification.
5. The method of claim 1 , further comprising detecting the presence of the target nucleic acid.
6. A method of amplifying a target nucleic acid sequence, comprising:
providing a split primer, wherein the split primer has two regions comprising a first sequence complementary to a first region of a target nucleic acid sequence and a second sequence complementary to a second region of a target nucleic acid, wherein the two regions of the split primer are not separated by an intervening base, and wherein the first and second regions of the target nucleic acid sequence are separated by at least one nucleic acid base;
hybridizing the split primer to the first and second regions of the target nucleic acid sequence in the presence of a polymerase and a reverse primer under conditions necessary for amplification; and
amplifying the target nucleic acid sequence.