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How To Calculate FST: A Step-by-Step Guide
  • Nov 15, 2024

How to Calculate FST: A Step-by-Step Guide

Fst, or Fixation index, is a widely used measure of genetic differentiation between populations. It is based on the concept of allele frequency, which refers to the proportion of a particular gene variant (allele) in a population. Fst measures the extent to which these allele frequencies differ between populations, and provides a quantitative estimate of the genetic structure of a species.



Calculating Fst requires genetic data from multiple populations, which can be obtained through various methods such as DNA sequencing or genotyping. Once the data is obtained, Fst can be calculated using different mathematical formulas, such as the Weir and Cockerham method or the Hudson estimator. The resulting Fst value ranges from 0 (no genetic differentiation) to 1 (complete genetic differentiation) and can be used to infer evolutionary processes such as migration, genetic drift, and natural selection.


Understanding how to calculate Fst is important for many fields of biology, including population genetics, evolutionary biology, and conservation genetics. By quantifying the genetic diversity and differentiation of populations, Fst can provide valuable insights into the evolutionary history and current status of a species.

Fundamentals of FST



Definition of FST


FST, also known as the fixation index, is a statistical measure that quantifies the degree of genetic differentiation between two or more populations. It was first introduced by Sewall Wright in 1931 as one of the components of his F-statistics. FST ranges from 0 to 1, where 0 indicates no genetic differentiation between populations, and 1 indicates complete differentiation.


FST is calculated by comparing the genetic variation within populations to the genetic variation between populations. The formula for FST is (HT - HS) / HT, where HT is the total genetic variation in the entire population, and HS is the average genetic variation within subpopulations.


Importance of FST in Population Genetics


FST is an important tool in population genetics because it allows researchers to quantify the degree of genetic differentiation between populations. This information can be used to understand the evolutionary history of populations, as well as to inform conservation and management strategies for endangered species.


FST can also be used to test hypotheses about the genetic structure of populations. For example, if two populations have a high FST value, it suggests that there is limited gene flow between them, which may be due to physical barriers or other factors that prevent individuals from moving between the populations.


Overall, FST is a powerful tool for understanding the genetic structure of populations and can provide valuable insights into the evolutionary history and conservation of species.

Calculating FST



FST is a measure of population differentiation due to genetic structure. It is frequently estimated from genetic polymorphism data, such as single-nucleotide polymorphisms (SNP) or microsatellites [1]. There are two commonly used methods to calculate FST: the allele frequency method and the variance components method.


Data Requirements


Before calculating FST, the following data are required:



  • Allele frequencies of each subpopulation

  • Total number of individuals in each subpopulation

  • Total number of alleles in each subpopulation


The Allele Frequency Method


The allele frequency method is the most commonly used method to calculate FST. It is based on the differences in allele frequencies between subpopulations [2]. The formula for calculating FST using the allele frequency method is:


FST = (HT - HS) / HT


Where HT is the total genetic diversity of the entire population, and HS is the average genetic diversity within subpopulations. The values of HT and HS can be calculated using the following formulas:


HT = (p)(1-p)


HS = (1/n) * Σpi(1-pi)


Where p is the frequency of the ith allele in the entire population, pi is the frequency of the ith allele in the ith subpopulation, and n is the number of subpopulations.


The Variance Components Method


The variance components method is an alternative method to calculate FST. It is based on the analysis of variance (ANOVA) of genetic variation within and between subpopulations [3]. The formula for calculating FST using the variance components method is:


FST = (σb^2) / (σb^2 + σw^2)


Where σb^2 is the variance between subpopulations, and σw^2 is the variance within subpopulations. The values of σb^2 and σw^2 can be calculated using the following formulas:


σb^2 = MSB - MSW


σw^2 = MSW


Where MSB is the mean square between subpopulations, and MSW is the mean square within subpopulations.


In conclusion, FST is a measure of population differentiation due to genetic structure. The allele frequency method and the variance components method are the two commonly used methods to calculate FST. The allele frequency method is based on the differences in allele frequencies between subpopulations, while the variance components method is based on the analysis of variance of genetic variation within and between subpopulations.

Interpreting FST Values



Range of FST


FST values range from 0 to 1, with 0 indicating no genetic differentiation between populations and 1 indicating complete genetic differentiation. An FST value of 0.05 or less is considered low, 0.05-0.15 is moderate, and greater than 0.15 is high.


Low, Moderate, and High FST Interpretations


When FST values are low, it suggests that populations are not genetically differentiated and are likely exchanging genes. In contrast, a high FST value suggests that populations are genetically isolated from one another and are not exchanging genes.


Moderate FST values indicate some degree of genetic differentiation, lump sum loan payoff calculator but not enough to suggest complete isolation. It is important to note that the interpretation of FST values depends on the context and the specific populations being studied.


Researchers should consider factors such as the natural history of the species, the geographic distance between populations, and the genetic markers used to estimate FST values. Additionally, FST values should be interpreted in conjunction with other analyses such as principal component analysis or phylogenetic analysis to gain a more complete understanding of the genetic relationships between populations.

Applications of FST



Evolutionary Biology


FST can be used to measure the degree of genetic differentiation between populations and can provide insights into the evolutionary history of a species. For example, FST values can be used to infer the level of gene flow between populations, which can help determine the extent to which populations have been isolated from each other. FST can also be used to estimate the time since populations diverged from a common ancestor, as well as the effective population size of each population.


Conservation Genetics


FST can be used to assess the genetic diversity of populations and to identify populations that are at risk of extinction. By comparing FST values between populations, conservation biologists can determine which populations are most genetically distinct and therefore most important for conservation efforts. FST can also be used to determine the degree of inbreeding within populations, which can be an important factor in conservation management.


Anthropology


FST can be used in anthropological studies to investigate patterns of human migration and population history. For example, FST values can be used to determine the degree of genetic differentiation between different human populations, which can provide insights into the history of human migration and the relationships between different populations. FST can also be used to investigate the genetic basis of human traits and diseases, such as lactose intolerance, which is associated with a specific allele at the LCT gene [1].


Overall, FST is a powerful tool for investigating genetic diversity and population structure, and can be applied to a wide range of fields, from evolutionary biology to conservation genetics and anthropology.


[1] Understanding FST - the fixation index - GitHub Pages

Challenges and Considerations



Sampling Error


One of the main challenges when calculating FST is sampling error. FST estimates are based on a sample of individuals from each population, and the smaller the sample size, the more likely it is that the estimate will be affected by sampling error. To reduce the impact of sampling error, researchers often use larger sample sizes, and some methods for calculating FST take into account the variability introduced by sampling error.


Mutation Rate


Another important consideration when calculating FST is the mutation rate. FST is based on the assumption that different populations have different allele frequencies due to genetic drift and/or natural selection, but not due to mutation. However, mutations can occur randomly and at different rates in different populations, which can lead to differences in allele frequencies that are not due to genetic drift or natural selection. This can result in an overestimation of FST if the mutation rate is not taken into account.


To address this issue, researchers often use methods that take into account the mutation rate, such as the method developed by Hudson et al. (1992) that uses a coalescent-based approach to estimate FST while accounting for mutation. However, it is important to note that the mutation rate can vary depending on the type of genetic marker used (e.g. microsatellites vs. SNPs), the genomic region being analyzed, and the species being studied.


Overall, it is important to carefully consider the potential sources of bias and error when calculating FST, and to use appropriate methods that take into account these factors. By doing so, researchers can obtain more accurate estimates of population differentiation and better understand the evolutionary processes shaping genetic diversity.

Frequently Asked Questions


What is the formula for calculating the fixation index (Fst)?


The formula for calculating Fst is based on the variance of allele frequencies within and between populations. The Fst value ranges from 0 to 1, where 0 indicates no genetic differentiation between populations and 1 indicates complete genetic differentiation. The formula for Fst is:


Fst = (Ht - Hs) / Ht


Where Ht is the total heterozygosity across all populations and Hs is the average heterozygosity within each population.


How can Fst values be interpreted in population genetics?


Fst values can be used to measure the degree of genetic differentiation between populations. Higher Fst values indicate greater genetic divergence between populations, while lower values indicate greater genetic similarity. Fst values can be used to infer historical patterns of migration, gene flow, and genetic drift within and between populations.


What steps are involved in measuring Fst between two populations?


Measuring Fst between two populations involves several steps, including: collecting genetic data, calculating allele frequencies, estimating heterozygosity, calculating Fst, and interpreting the results. The choice of genetic markers, sample size, and statistical methods can all affect the accuracy and precision of Fst estimates.


How can allele frequencies be used to calculate Fst?


Allele frequencies can be used to calculate Fst by comparing the variance of allele frequencies within and between populations. Fst can be calculated using different methods, including the Weir and Cockerham method, the Nei method, and the AMOVA method. These methods differ in their assumptions about the underlying genetic model and the level of population structure.


What methods are available for calculating Fst in R programming language?


Several R packages are available for calculating Fst, including hierfstat, adegenet, and poppr. These packages implement various methods for estimating Fst, including the Weir and Cockerham method, the AMOVA method, and the pairwise method. R provides a flexible and powerful environment for analyzing genetic data and exploring patterns of population structure.


How does the Hardy-Weinberg equilibrium relate to Fst calculations?


The Hardy-Weinberg equilibrium (HWE) is a fundamental principle in population genetics that describes the relationship between allele frequencies and genotype frequencies in a population. Fst calculations assume that populations are in HWE, meaning that random mating and genetic drift are the only forces affecting allele frequencies. Deviations from HWE can lead to biased Fst estimates and should be taken into account when interpreting the results.

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