The present teachings generally relate to methods and kits incorporating a discriminating positive control for determining whether a particular microorganism or group of microorganisms are present in a sample, for example but not limited to a food, environmental, agricultural, biopharmaceutical, pharmaceutical, or water sample. According to certain methods, at least part of a starting material, for example but not limited to, a food, environmental, agricultural, biopharmaceutical, pharmaceutical, or water sample can be combined with a culture medium and incubated under conditions suitable for microbial growth followed by extracting microorganism and added control nucleic acids for analysis. The extracted nucleic acids are amplified and the amplified nucleic acids are detected, directly or indirectly, and the fidelity of the methods and the presence or absence of the corresponding microorganism is determined because the discriminating positive control provides both confirmatory results for the methods used but eliminates false positive results.

The present teachings generally relate to methods and kits incorporating a discriminating positive control for determining whether a particular microorganism or group of microorganisms are present in a sample, for example but not limited to a food, environmental, agricultural, biopharmaceutical, pharmaceutical, or water sample. According to certain methods, at least part of a starting material, for example but not limited to, a food, environmental, agricultural, biopharmaceutical, pharmaceutical, or water sample can be combined with a culture medium and incubated under conditions suitable for microbial growth followed by extracting microorganism and added control nucleic acids for analysis. The extracted nucleic acids are amplified and the amplified nucleic acids are detected, directly or indirectly, and the fidelity of the methods and the presence or absence of the corresponding microorganism is determined because the discriminating positive control provides both confirmatory results for the methods used but eliminates false positive results.

Methods and compositions are provided for quantifying the number of biological input molecules of one or more target genes of interest in a PCR. The method comprises steps for amplifying multiple synthetic templates and high throughput sequencing. The method further comprises steps for achieving equal sequencing coverage between targets that do not occur in equal ratios, while mitigating the need for costly deep sequencing methods.

Methods and compositions are provided for quantifying the number of biological input molecules of one or more target genes of interest in a PCR. The method comprises steps for amplifying multiple synthetic templates and high throughput sequencing. The method further comprises steps for achieving equal sequencing coverage between targets that do not occur in equal ratios, while mitigating the need for costly deep sequencing methods.

What is described is a method for quantitative and qualitative analysis of complex template nucleic acids to be analyzed. The method comprises the co-amplification of a control nucleic acid with the complex template nucleic acid by means of isothermal strand displacement reaction. The method of the invention further comprises the determination of the amount of amplified control nucleic acid as measure for the determination of the quantity and/or quality of the complex template nucleic acid used. The present invention also relates to a kit for carrying out a method of the invention. Furthermore, the use of the method of the invention or the kit of the invention for standardization of whole-genome, whole-transcriptome and whole-bisulfitome analyses is described.

What is described is a method for quantitative and qualitative analysis of complex template nucleic acids to be analyzed. The method comprises the co-amplification of a control nucleic acid with the complex template nucleic acid by means of isothermal strand displacement reaction. The method of the invention further comprises the determination of the amount of amplified control nucleic acid as measure for the determination of the quantity and/or quality of the complex template nucleic acid used. The present invention also relates to a kit for carrying out a method of the invention. Furthermore, the use of the method of the invention or the kit of the invention for standardization of whole-genome, whole-transcriptome and whole-bisulfitome analyses is described.

Methods are provided for determining the methylation status of GC-rich templates. The methods include use of GC reference standards that allow simultaneous characterization of methylation status and CGG repeat length. The methods are useful for detecting genotypes associated with GC-rich repeats, including Fragile X Syndrome.

Methods are provided for determining the methylation status of GC-rich templates. The methods include use of GC reference standards that allow simultaneous characterization of methylation status and CGG repeat length. The methods are useful for detecting genotypes associated with GC-rich repeats, including Fragile X Syndrome.

Method of analysis for alleles of a target. In the method, a plurality of fluid volumes may be formed. Each fluid volume may contain a first primer pair to amplify a first allele of a target, a second primer pair to amplify a second allele of the target, a first reporter including a first photoluminophore and providing a primer of the first primer pair, and a second reporter including a second photoluminophore and providing a primer of the second primer pair. Each of the first and second reporters may include an oligomer having a quencher, and the oligomer may be configured to base-pair with the primer of the first primer pair and the primer of the second primer pair. The first and second alleles may be amplified using the first and second primer pairs. Photoluminescence may be detected. A level of each allele may be determined.

Method of analysis for alleles of a target. In the method, a plurality of fluid volumes may be formed. Each fluid volume may contain a first primer pair to amplify a first allele of a target, a second primer pair to amplify a second allele of the target, a first reporter including a first photoluminophore and providing a primer of the first primer pair, and a second reporter including a second photoluminophore and providing a primer of the second primer pair. Each of the first and second reporters may include an oligomer having a quencher, and the oligomer may be configured to base-pair with the primer of the first primer pair and the primer of the second primer pair. The first and second alleles may be amplified using the first and second primer pairs. Photoluminescence may be detected. A level of each allele may be determined.