Abstract: The invention relates to a method for single cell analysis of relative telomere length using multiplex pre-PCR followed by a qPCR (SCT-pqPCR).

Abstract: The invention relates to a method for single cell analysis of relative telomere length using multiplex pre-PCR followed by a qPCR (SCT-pqPCR).

Abstract: The present invention includes a method to identify stem cell genes that are differentially expressed in stem cells at various stages of differentiation when compared to undifferentiated stem cells by preparing a gene expression profile of a stem cell population and comparing the profile to a profile prepared from stem cells at different stages of differentiation, thereby identifying cDNA species, and therefore genes, which are expressed. The present invention also includes methods to identify a therapeutic agent that modulates the expression of at least one stem cell gene associated with the differentiation, proliferation and/or survival of stem cells.

Abstract: Methods are disclosed to identify T lymphocyte genes that are differentially expressed upon exposure to a pathogen (viral or bacterial), immunogen, antigen, or in a sterile inflammatory disease, autoimmune disease, immunodeficiency disease, lymphocytic cancers, or graft versus host rejection. The method involves the preparation of a gene expression profile of a T lymphocyte population exposed to a pathogen or isolated from a subject having one of the aforementioned pathologies and comparing that profile to a profile prepared from quiescent T lymphocytes. The present invention is useful for identifying cytokine genes, genes encoding cell surface receptors and genes encoding intermediary signalling molecules. Related methods for identifying therapeutic or prophylatic immunomodulatory agents are present.

Abstract: An improved method for screening genomic, cDNA, or any DNA fragments in general is described. Novel adapters are ligated to the ends of DNA fragments from two different individuals or two different pools of individuals. The DNA fragment-adapter complexes are mixed, denatured and reannealed to form homohybrid and heterohybrid DNA duplexes, which are separated based on characteristics of the adapters. Sequence differences between heterohybrids can be revealed as mismatched base pairs in the heterohybrid DNA duplex. Mismatch base pairs are discovered using genome mismatch scanning techniques that use thymine glycosylases and related enzymes that capture DNA containing mismatched base pairs. The perfectly base paired DNA or DNA containing mismatched base pairs can be further separated into homohybrids and heterohybrids using novel adapters that allow physical capture of either heterohybrid or homohybrid DNA.

Abstract: The present invention comprises a method to identify granulocytic cell genes that are differentially expressed upon exposure to a pathogen or in a sterile inflammatory disease by preparing a gene expression profile of a granulocytic cell population exposed to a pathogen or isolated from a subject having a sterile inflammatory disease and comparing that profile to a profile prepared from quiescent granulocytic cells. The present invention is particularly useful for identifying cytokine genes, genes encoding cell surface receptors and genes encoding intermediary signaling molecules. The invention also includes methods to identify a therapeutic agent that modulates the expression of at least one gene in a granulocyte population. Genes which are differentially expressed during neutrophil contact with a pathogen, such as a virulent bacteria, or that are differentially expressed in a subject having a sterile inflammatory disease are of particular importance.

Abstract: DNA containing nucleotide base mispairs can be isolated using a modified rolling circle amplification procedure. Specific Y-shaped adapters permit the selective circularization of these fragments with a complementary splint oligonucleotide. Rolling circle amplification is then carried out with a DNA polymerase. Rolling circle amplification can also be carried out using a mixture of DNA circles having different lengths. Genetic phase of linked DNA markers can be determined by selective amplification of one parental haplotype. DNA fragments can also be converted into a form that can be utilized as rolling circle amplification templates by ligation of hairpin forming adapters to the ends of the fragments. Two or more DNA polymerases can be used in a rolling circle amplification reaction. DNA polymerase III has special properties that improve rolling circle amplification.

Abstract: DNA containing nucleotide base mispairs can be isolated using a modified rolling circle amplification procedure. Specific Y-shaped adapters permit the selective circularization of these fragments with a complementary splint oligonucleotide. Rolling circle amplification is then carried out with a DNA polymerase. Rolling circle amplification can also be carried out using a mixture of DNA circles having different lengths. Genetic phase of linked DNA markers can be determined by selective amplification of one parental haplotype. DNA fragments can also be converted into a form that can be utilized as rolling circle amplification templates by ligation of hairpin forming adapters to the ends of the fragments. Two or more DNA polymerases can be used in a rolling circle amplification reaction. DNA polymerase III has special properties that improve rolling circle amplification.

Abstract: Genomic or cDNA, or fragments and mixtures thereof, can be screened by generation of subsets and then subjecting the subsets to mismatch scanning procedures. Alternatively, DNA fragments can be generated by cutting with a restriction endonuclease that generates variable overhangs. For either of the above methods, Y-shaped adapters having a region of non-complementary single-stranded DNA at the end can be used. Heterohybrid DNA, containing one DNA strand derived from each of two different samples, or homohybrids, containing DNA strands from the same sample, can be selected. Adapters attached to the ends of the fragments are designed to allow the selective isolation of homohybrid or heterohybrid DNA.

Abstract: Genomic or cDNA, or fragments and mixtures thereof, can be screened by generation of subsets and then subjecting the subsets to mismatch scanning procedures. Alternatively, DNA fragments can be generated by cutting with a restriction endonuclease that generates variable overhangs. For either of the above methods, Y-shaped adapters having a region of non-complementary single-stranded DNA at the end can be used. Heterohybrid DNA, containing one DNA strand derived from each of two different samples, or homohybrids, containing DNA strands from the same sample, can be selected. Adapters attached to the ends of the fragments are designed to allow the selective isolation of homohybrid or heterohybrid DNA.

Abstract: DNA containing nucleotide base mispairs can be isolated using a modified rolling circle amplification procedure. Specific Y-shaped adapters permit the selective circularization of these fragments with a complementary splint oligonucleotide. Rolling circle amplification is then carried out with a DNA polymerase. Rolling circle amplification can also be carried out using a mixture of DNA circles having different lengths. Genetic phase of linked DNA markers can be determined by selective amplification of one parental haplotype. DNA fragments can also be converted into a form that can be utilized as rolling circle amplification templates by ligation of hairpin forming adapters to the ends of the fragments. Two or more DNA polymerases can be used in a rolling circle amplification reaction. DNA polymerase III has special properties that improve rolling circle amplification.

Abstract: Genomic or cDNA, or fragments and mixtures thereof, can be screened by generation of subsets and then subjecting the subsets to mismatch scanning procedures. Alternatively, DNA fragments can be generated by cutting with a restriction endonuclease that generates variable overhangs. For either of the above methods, Y-shaped adapters having a region of non-complementary single-stranded DNA at the end can be used. Heterohybrid DNA, containing one DNA strand derived from each of two different samples, or homohybrids, containing DNA strands from the same sample, can be selected. Adapters attached to the ends of the fragments are designed to allow the selective isolation of homohybrid or heterohybrid DNA.

Abstract: DNA containing nucleotide base mispairs can be isolated using a modified rolling circle amplification procedure. Specific Y-shaped adapters permit the selective circularization of these fragments with a complementary splint oligonucleotide. Rolling circle amplification is then carried out with a DNA polymerase. Rolling circle amplification can also be carried out using a mixture of DNA circles having different lengths. Genetic phase of linked DNA markers can be determined by selective amplification of one parental haplotype. DNA fragments can also be converted into a form that can be utilized as rolling circle amplification templates by ligation of hairpin forming adapters to the ends of the fragments. Two or more DNA polymerases can be used in a rolling circle amplification reaction. DNA polymerase III has special properties that improve rolling circle amplification.

Abstract: A general method for screening genomic or cDNA, or fragments and mixtures thereof, involves sample simplification by the generation of subsets and then subjecting the subsets to a modified mismatch scanning procedure that eliminates DNA having single stranded breaks after a MutSLH cleavage. The methods are particularly useful in human population isolates, including identification of identical-by-descent sequences, genomic comparisons of two or more individuals, and genomic comparisons of two populations of individuals, for the identification of sequences of low polymorphism.

Abstract: A stable, single-stranded nucleoprotein filament adapted to complex specifically and stably with a target duplex DNA having a selected base sequence. The filament is composed of a single-stranded DNA probe having a region of homology with the target base sequence, and RecA protein bound stably to the DNA probe by adenosine 5'-(.gamma.-thio)triphosphate. The filament is useful in a novel system and method for enriching target duplex DNA which contains a region homologous to the probe sequence, for blocking selected restriction endonuclease sites in the target DNA, and in other DNA methodologies in which stable rapid triple-strand synaptic formation in duplex DNA can be exploited.