Method of pooling samples for performing a bi0l0gical assay

A sample, in-sample technique, applied in the field of measurement with classification results, to achieve the effect of reducing the cost of analysis

Inactive Publication Date: 2010-12-08
HENDRIX GENETICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0014] An important disadvantage of sample pooling is that the property being measured is only determined in the pool as a whole, not in any individual sample in that pool

Method used

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  • Method of pooling samples for performing a bi0l0gical assay
  • Method of pooling samples for performing a bi0l0gical assay
  • Method of pooling samples for performing a bi0l0gical assay

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] Example of genotyping a sample of diploid individuals for the presence of a SNP using a normalized pool of 50 individuals

[0097] Step 1) Test 50 individuals individually

[0098] For each SNP and each individual, we obtained the intensities of red fluorescence (allele present) and green fluorescence (allele absent) in microarray format using two different fluorescent dyes. The ratio between red and green intensities is not always 1 (or 0) for animals that are homozygous or 0.5 for animals that are heterozygous.

[0099] Individual genotyped data were used to calculate correction factors for the signal intensities of all genotyped SNPs.

[0100] To obtain the most important correction factor (K), which is usually used to correct data representing arbitrary unequal efficiencies among alleles, we use the signal from the heterozygous genotype. If no heterozygous genotypes are present, we assume that the SNP under study does not segregate in the population under study, a...

Embodiment 2

[0141] Example of Genotyping Diploid Individual Samples Using a Pool of 25 Normalized 2 Individuals

[0142] Step 1) As in Step 1 of Example 1, 50 individuals were individually tested.

[0143] Step 2) Construct 25 pools at a ratio of 1:3, with 2 samples in each pool, which include all 50 individuals from step 1) above. In these pools, the allele frequencies are estimated uncorrected or based on the correction factors obtained in the first step.

[0144] Step 3) Compare the sum of the typed allele frequencies of the 2 individuals to the estimated frequency in the pool with a sample of 2 individuals. Compute the regression line from these 25 points. The regression coefficients and intercepts can then be used to correct the estimated frequencies for the other pools.

[0145] Step 4) Then at 1:3, 1:3:9 or 1:3 1 : 3 2 : 3 (n-1) Construct DNA pools of 2, 3 or n samples at a ratio of .

[0146] Step 5) With the correction factors obtained in steps 1 and 3, calculate the allel...

Embodiment 3

[0149] Example of genotyping samples from haploid individuals

[0150] When pooling two haploid samples and measuring the presence of allele A at certain locations in the genome, the ratios in the expected measurements (peak height, surface area, intensity) are;

[0151] Table 4. Results points for allele frequencies of pooled samples and deduced genotypes for two individuals in the pool of SNPs with A and C alleles

[0152] Allelic of pooled samples

frequency of gene A

Deduced genotype of individual 1

(takes 1 share in the pool)

Deduced genotype of individual 2

(takes 3 shares in the pool)

0.00

C

C

0.33

A

C

0.67

C

A

[0153] 1.00

A

A

[0154] If only two-sample pools are used, no correction factor may be required. Correction factors may be required when more samples are pooled. It can then be calculated by simulating a pool of 2 samples of heterozygous and hom...

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Abstract

The present invention relates to a method of pooling samples to be analyzed for a categorical variable, wherein the analysis involves a quantitative measurement of an analyte, said method of pooling samples comprising providing a pool of n samples wherein the amount of individual samples in the pool is such that the analytes in the samples are present in a molar ratio of x0 : x1 : x2 : x(n-1), and wherein x is an integer of 2 or higher representing the number of classes of the categorical variable.

Description

technical field [0001] The present invention relates to the field of measurements with classified results on biological samples, and more particularly to a sample preparation method for biological assays with classified results. The invention provides a method of pooling samples and the use of said method in genotyping of allelic variables. The invention also provides a method of analyzing a plurality of samples, a combining device for combining a plurality of samples into a pooled sample, an analysis device comprising a processor for analyzing a series of pooled samples, a computer program for implementing a method of pooling samples Products and computer program products implementing methods for analyzing multiple samples. Background technique [0002] A bioassay is a method of determining the identity, concentration or presence of a biological analyte in a sample. Bioassays are an inherent part of research in all fields of science, most notably in the life sciences, esp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6874C12Q1/6827C12Q1/6806C12Q2537/143C12Q2563/107
Inventor 阿德里安乌斯·拉姆贝图斯·约翰纳斯·韦雷吉肯安内米克·波拉·容格乌斯赫拉尔杜斯·安东尼厄斯·阿诺尔德斯·阿尔贝斯
Owner HENDRIX GENETICS
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