Anchored strand displacement amplification on an electronically addressable microchip
First Claim
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1. A strand displacement amplification primer set comprising:
- a. at least one pair of single stranded nucleic acid sequences, a first portion of said sequences having sequence complementary with a target nucleic acid of interest, the portion of said single stranded nucleic acid sequences of said pair having sequence complementary with a target nucleic acid of interest further having complementary sequence to said target nucleic acid wherein one of said pair is complementary to a plus strand of said target and the other of said pair is complementary to a minus strand of said target;
b. a second portion of said sequences encoding either a plus or a minus strand of at least one endonuclease restriction site; and
c. a molecular structure having covalent attachment moieties for binding a 5′
terminus of each single stranded nucleic acid sequence of said pair either covalently or non-covalently to a solid support.
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Abstract
Methods and compositions of matter are provided for the strand displacement amplification of target nucleic acids of interest using primer pair sets that are anchored to electronically addressable capture sites on a microarray. The primer pair sets may be individually bound to the capture sites or may comprise a unique branched primer pair moiety. The anchored primers allow for the simultaneous multiplex capture, amplification and detection of a target nucleic acid derived from any sample source.
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12 Claims
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1. A strand displacement amplification primer set comprising:
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a. at least one pair of single stranded nucleic acid sequences, a first portion of said sequences having sequence complementary with a target nucleic acid of interest, the portion of said single stranded nucleic acid sequences of said pair having sequence complementary with a target nucleic acid of interest further having complementary sequence to said target nucleic acid wherein one of said pair is complementary to a plus strand of said target and the other of said pair is complementary to a minus strand of said target;
b. a second portion of said sequences encoding either a plus or a minus strand of at least one endonuclease restriction site; and
c. a molecular structure having covalent attachment moieties for binding a 5′
terminus of each single stranded nucleic acid sequence of said pair either covalently or non-covalently to a solid support. - View Dependent Claims (2, 3, 4, 5)
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6. A branched strand displacement amplification primer set comprising:
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a. at least one pair of single stranded nucleic acid sequences, a first portion of said sequences having sequence complementary with a target nucleic acid of interest, the portion of said single stranded nucleic acid sequences of said pair having sequence complementary with a target nucleic acid of interest further having complementary sequence to said target nucleic acid wherein one of said pair is complementary to a plus strand of said target and the other of said pair is complementary to a minus strand of said target;
b. a second portion of said sequences encoding either a plus or a minus strand of at least one endonuclease restriction site; and
c. a branched molecular structure having covalent attachment moieties for binding a 5′
terminus of each single stranded nucleic acid sequence of said pair and attachments for binding either covalently or non-covalently to a solid support. - View Dependent Claims (7, 8, 9, 10)
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11. A kit for carrying out anchored SDA reactions for use on a bioelectronic microchip comprising:
a. one or more oligonucleotides for use on a bioelectronic microchip specific for Factor V, Hemochromotosis, or a bacterium, which oligonucleotides comprise amplification primers, bumper primers, capture probes, and/or signal probes selected from the group consisting of Seq. Id. Nos. 20, 21, 31, 32, 45, 46, 51, 52, 57, and 58. - View Dependent Claims (12)
Specification