Bio-rad Laboratories, Inc.

City of Hope

Luis A Ugozzoli

R Bruce Wallace

Robert Bruce Wallace

Claudia Litterst

Bijay K Pal

Camille M Diges

Hector Juarez-Salinas

Antonio Arevalo Reyes

Dan Y Wu

A method of characterizing telomere length. A template may be synthesized in a bulk phase reaction mixture containing genomic DNA. Partitions may be formed after synthesizing the template in the bulk phase reaction mixture, with only a subset of the partitions containing at least one copy of the template. At least a region of the template may be amplified in partitions. Amplification data may be collected from partitions. A measure of telomere length may be determined for the genomic DNA based on the amplification data. In some embodiments, the bulk phase reaction mixture may be thermally cycled for at least two cycles, and heated to a denaturation temperature and cooled to an annealing temperature in each cycle.

Digital assay for telomere length

RNA is extracted from cellular material with a reagent that includes heparin, a reducing agent to reduce disulfide bonds, a chelating agent, a buffer, and an alkali metal halide. The reagent does not require the use of organic solvents, and the reagent allows extraction to be performed in a relatively short period of time in comparison to the prior art.

Cell lysis reagent for isolation of RNA

The extensive synthesis (&ldquo;amplification&rdquo;) of a nucleic acid sequence of interest is attained through a linked series of multi-cycle primer extension reactions (LLA). The primers used in each of the primer extension reactions of the process contain non-replicable and/or cleavable elements that halt nucleic acid synthesis and thereby prevent the synthesized molecules from serving as templates in subsequent cycles. Synthesized molecules accumulate during primer extension in a mathematically linear fashion, thereby rendering the process relatively insensitive to contaminating nucleic acids. Multiple primer sets are employed, and simultaneously included in the reaction mixture, thereby ensuring the accumulation of a large number of copies of the nucleic acid sequence of interest. The invention also provides for the detection of an amplified nucleic acid sequence of interest, as well as reagent kits for carrying out the reaction.

Linked linear amplification of nucleic acids

A rapid, non-radioactive approach to the diagnosis of sickle cell anemia is described based on an allele specific polymerase chain reaction (ASPCR) in which the 3'-terminal nucleotide of one of the primers of the primer set forms a match with one allele and a mismatch with the other allele. This method allows the direct detection of the normal or the sickle cell .beta.-globin allele in genomic DNA without the additional steps of probe hybridization, ligation or restriction enzyme cleavage.

Allele specific polymerase chain reaction

Disclosed is an application of allele specific polymerase chain reaction technology for the direct determination of multiple allele genotyping. ABO genotyping is demonstrated with allele specific primer sets.

Genotyping of multiple allele systems

This invention provides methods and compounds relating to the use of deoxyribose nicotinamide adenine dinucleotide (dNAD.sup.+) analogues in NAD.sup.+ -dependent ligation reactions. The compounds and methods may be used in NAD.sup.+ -dependent ligation reactions generally, and to increase the specificity of NAD.sup.+ -dependent ligation reactions.

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San Rafael, CA, United States of America

Luis A Ugozzoli