Methods for using riboprimers for strand displacement replication of target sequences
First Claim
Patent Images
1. A method for amplifying a target nucleic acid sequence comprising a target nucleic acid:
- a) hybridizing a Riboprimer to a single stranded DNA template comprising the target nucleic acid sequence, wherein said Riboprimer comprises;
i) only ribonucleotides, or ii) only purine ribonucleotides and only pyrimidine nucleotides, wherein at least one of the pyrimidine nucleotides is a pyrimidine 2′
-deoxyribonucleotide having a non-canonical substituent, which substituent is neither an H nor an OH, on the 2′
-position of the deoxyribose sugar moiety;
b) extending the Riboprimer with a DNA polymerase having strand displacement activity;
c) cleaving the annealed Riboprimer with an RNase H enzyme such that another Riboprimer hybridizes to the template and repeats primer extension by strand displacement, whereby multiple copies of the complementary sequence of the target sequence are produced; and
d) attaching the multiple copies of the complementary sequence of the target sequence onto a solid substrate to make a microarray of the amplified products.
4 Assignments
0 Petitions
Accused Products
Abstract
Methods, compositions and kits for amplifying a target sequence by strand displacement replication using strand-displacing primers. The method uses primers that have only ribonucleotides or purine ribonucleotides and at least one 2′-substituted pyrimidine-2′-deoxyribonucleotide.
61 Citations
13 Claims
-
1. A method for amplifying a target nucleic acid sequence comprising a target nucleic acid:
-
a) hybridizing a Riboprimer to a single stranded DNA template comprising the target nucleic acid sequence, wherein said Riboprimer comprises;
i) only ribonucleotides, or ii) only purine ribonucleotides and only pyrimidine nucleotides, wherein at least one of the pyrimidine nucleotides is a pyrimidine 2′
-deoxyribonucleotide having a non-canonical substituent, which substituent is neither an H nor an OH, on the 2′
-position of the deoxyribose sugar moiety;b) extending the Riboprimer with a DNA polymerase having strand displacement activity; c) cleaving the annealed Riboprimer with an RNase H enzyme such that another Riboprimer hybridizes to the template and repeats primer extension by strand displacement, whereby multiple copies of the complementary sequence of the target sequence are produced; and d) attaching the multiple copies of the complementary sequence of the target sequence onto a solid substrate to make a microarray of the amplified products. - View Dependent Claims (2, 3, 4, 5, 6, 7, 11, 12, 13)
-
-
8. A method for amplifying a target nucleic acid sequence comprising a target nucleic acid:
-
a) hybridizing a Riboprimer to a single stranded DNA template comprising the target nucleic acid sequence, wherein said Riboprimer comprises;
i) only ribonucleotides, or ii) only purine ribonucleotides and only pyrimidine nucleotides, wherein at least one of the pyrimidine nucleotides is a pyrimidine 2′
-deoxyribonucleotide having a non-canonical substituent, which substituent is neither an H nor an OH, on the 2′
-position of the deoxyribose sugar moiety;b) extending the Riboprimer with a DNA polymerase having strand displacement activity; c) cleaving the annealed Riboprimer with an RNase H enzyme such that another Riboprimer hybridizes to the template and repeats primer extension by strand displacement, whereby multiple copies of the complementary sequence of the target sequence are produced; and d) hybridizing the multiple copies of the complementary sequence of the target sequence to a microarray of nucleic acid molecules immobilized on a surface of a solid phase. - View Dependent Claims (9, 10)
-
Specification
-
- Resources
Litigation Campaign Assessment
Thank you for your request. You will receive a custom alert email when the Litigation Campaign Assessment is available.
×
-
Current AssigneeIllumina Incorporated
-
Original AssigneeEpicentre Technologies Corporation (Illumina Incorporated)
-
InventorsDahl, Gary A., Jendrisak, Jerome J., Radek, Agnes J.
-
Primary Examiner(s)Chunduru; Suryaprabha
-
Application NumberUS12/186,127Publication NumberTime in Patent Office553 DaysField of Search435/6, 435/91.2, 536/24.31, 536/24.32, 536/24.33US Class Current435/6.12CPC Class CodesC12N 15/1096 cDNA Synthesis; Subtracted ...C12Q 1/6853 using modified primers or t...C12Q 2521/301 EndonucleaseC12Q 2525/125 incorporating agents result...C12Q 2531/119 Strand displacement amplifi...
