摘要:
Methods for determining the binding affinity of a probe to a target or targets in a polynucleotide composite using an automated system. Such methods determine relative binding sites based on thermodynamic principles, using a thermodynamic alphabet and a thermodynamic scoring matrix, with appropriate computer software, such as BLASTP. The invention also provides methods for designing polynucleotide probes to be used in hybridization assays, and minimizing the occurrence of cross-hybridization.
摘要:
Methods and systems for ordering assays which detect SNPs or gene expression are provided. The methods use PCR and RT-PCR procedures. Collections of stock assays are assembled using pre- and post-manufacturing quality control procedures and made available to consumers via the Internet. In addition, custom assays are prepared upon order from the consumer and these assays are also prepared using pre- and post-manufacturing quality control procedures. The assays are then delivered to the consumer.
摘要:
In some embodiments, the present teachings provide a method of identifying a nucleotide of interest in a target polynucleotide. In some embodiments, the method can comprise forming an amplification strand, wherein the amplification strand comprises a hairpinning end region; hybridizing the hairpinning end region of the amplification strand with a hairpinning region of a target polynucleotide, to form a self-complementary amplification product. After performing an extension reaction, wherein the hairpinning end region of the amplification strand is extended, an extended reaction product is formed. Detection of the extended reaction product can result in the identification of a nucleotide of interest in the target polynucleotide. Additional methods, as well as kits, are also provided.
摘要:
The present teachings provide methods, compositions, and kits for querying the identity of a target polynucleotide strand comprising. In some embodiments, the present teachings provide a method comprising forming a reaction complex comprising the target polynucleotide strand hybridized to an upstream probe, a middle probe, and a downstream probe, wherein the middle probe comprises, A) a first target specific portion, B) a second target specific portion, C) a non-target specific portion, wherein the non-target specific portion is located between the first target specific portion and the second target specific portion, wherein the downstream probe comprises a 5′ end that is adjacent with the 3′ end of the middle probe, wherein the upstream probe comprises a 3′ end that is adjacent with the 5′ end of the middle probe; ligating the upstream probe to the middle probe and the middle probe to the downstream probe to form a ligation product; detecting the ligation product; and, determining the identity of the target polynucleotide strand.
摘要:
The present teachings generally relate to methods, kits, and compositions for detecting target polynucleotide sequences. The teachings also relate to ligation and amplification reactions that generate self-complementary polynucleotide products. In some embodiments, ligation reactions are performed with probes that result in the formation a self-complementary ligation product. Ligation of a hairpin linker to the self-complementary ligation product can form a loop ligation product. In some embodiments, the loop ligation product can be amplified with rolling circle amplification. Detection of a loop ligation product can serve to determine the identity of a target polynucleotide.
摘要:
The present teachings generally relate to methods, kits, and compositions for detecting target polynucleotide sequences. The teachings also relate to ligation and amplification reactions that generate self-complementary polynucleotide products. In some embodiments, ligation reactions are performed with probes that result in the formation a self-complementary ligation product. Ligation of a hairpin linker to the self-complementary ligation product can form a loop ligation product. In some embodiments, the loop ligation product can be amplified with rolling circle amplification. Detection of a loop ligation product can serve to determine the identity of a target polynucleotide.
摘要:
The present invention relates to methods and kits for detecting the presence or absence of (or quantitating) target nucleic acid sequences using ligation and amplification. The invention also relates to methods, reagents, and kits that employ addressable portions and labeled probes.