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Determination of allele frequency from DNA pools using bovine trinucleotide microsatellites
Year:
1998
Source of publication :
Animal Biotechnology
Authors :
Ron, Micha
;
.
Volume :
9
Co-Authors:
Band, M., Institute of Animal Sciences, Volcani Center, P.O.B. 6, Bet Dagan, 50250, Israel
Ron, M., Institute of Animal Sciences, Volcani Center, P.O.B. 6, Bet Dagan, 50250, Israel
Facilitators :
From page:
35
To page:
45
(
Total pages:
11
)
Abstract:
Three sets of fluorescent labelled primers were used to amplify bovine trinucleotide microsatellites from DNA pools. DNA from 20 individuals were collected to create 3 pools differing in allele frequencies. Replicate mixes from each pool were used as template for PCR reactions. PCR products were separated and quantified on an automated DNA sequencer. Allele frequency estimates from pooled samples corrected for overlapping shadow peaks were calculated. Rare alleles representing only 2.5% of the total pool were accurately detected. Standard error of allele frequency estimates expressed as percent of the total 40 chromosomes per pool ranged between 0.8%-4.6% for different microsatellite-pool combinations as compared to 8.0% binomial sampling error. Regression coefficients of actual allele frequencies, determined by individual genotyping, on estimated frequencies ranged from 0.96-1.06. As regression slopes were close to unity it can be deduced that corrected peak height values from a DNA pool are unbiased estimates of actual allele frequencies. With standard error of the y-intercept of 0.21, the 95% confidence interval of allele frequency is 0.42 alleles or 1% in a pool of 40 chromosomes. Thus, it would be possible to detect an allele with a frequency of greater than 1% within the pool.
Note:
Related Files :
Alleles
Animal
animal disease
Animals
cattle
DNA
epidemiology
gene frequency
Genetics
Trinucleotide Repeats
Show More
Related Content
More details
DOI :
Article number:
Affiliations:
Database:
Scopus
Publication Type:
article
;
.
Language:
English
Editors' remarks:
ID:
30539
Last updated date:
02/03/2022 17:27
Creation date:
17/04/2018 00:55
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Scientific Publication
Determination of allele frequency from DNA pools using bovine trinucleotide microsatellites
9
Band, M., Institute of Animal Sciences, Volcani Center, P.O.B. 6, Bet Dagan, 50250, Israel
Ron, M., Institute of Animal Sciences, Volcani Center, P.O.B. 6, Bet Dagan, 50250, Israel
Determination of allele frequency from DNA pools using bovine trinucleotide microsatellites
Three sets of fluorescent labelled primers were used to amplify bovine trinucleotide microsatellites from DNA pools. DNA from 20 individuals were collected to create 3 pools differing in allele frequencies. Replicate mixes from each pool were used as template for PCR reactions. PCR products were separated and quantified on an automated DNA sequencer. Allele frequency estimates from pooled samples corrected for overlapping shadow peaks were calculated. Rare alleles representing only 2.5% of the total pool were accurately detected. Standard error of allele frequency estimates expressed as percent of the total 40 chromosomes per pool ranged between 0.8%-4.6% for different microsatellite-pool combinations as compared to 8.0% binomial sampling error. Regression coefficients of actual allele frequencies, determined by individual genotyping, on estimated frequencies ranged from 0.96-1.06. As regression slopes were close to unity it can be deduced that corrected peak height values from a DNA pool are unbiased estimates of actual allele frequencies. With standard error of the y-intercept of 0.21, the 95% confidence interval of allele frequency is 0.42 alleles or 1% in a pool of 40 chromosomes. Thus, it would be possible to detect an allele with a frequency of greater than 1% within the pool.
Scientific Publication
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