# The Huntington Library, Art Collections, and Botanical Gardens Bean There, Done That: A Hardy-Weinberg Simulation

This lesson plan was adapted from San Francisco State University’s Bio 240: http://userwww.sfsu.edu/~biol240/labs/lab_02hardyweinberg/pages/lab_01expt.html

Overview Students will look at a population of beans to examine conditions under which the Hardy-Weinberg equilibrium applies. Introduction In the first part of this exercise we will demonstrate the contention of the HardyWeinberg Theorem that genetic recombination as a result of sexual reproduction will not, by itself, cause any change in allele frequencies or genotype frequencies in a population from one generation to the next. Obviously, it is difficult to demonstrate this principal using real organisms: depending on the organism, it could take days, weeks, months or even years to show that allele and genotype frequencies remained constant for several generations - or even for a single generation. Furthermore, the Hardy-Weinberg equilibrium depends on the absence of any forces operating which might change allele frequencies in a population. These forces—mutation, migration, selection, and chance effects—are almost never absent from natural populations and it would be very difficult even in the laboratory to erect a set of conditions that might reliably exclude them. The Hardy-Weinberg Law can be mathematically demonstrated in the following table. If p equals the frequency of allele A and q is the frequency of allele a, union of gametes would occur as follows: p q p p2 pq q pq q2 One of the predictions of the Hardy-Weinberg Law refers to the genotypic frequencies after one generation of random mating. In the above table, the genotypic frequencies for AA is p2, the genotypic frequency for Aa is 2pq and the genotypic frequency for aa will be q2. These are the values that are predicted by the law. The second prediction is that the frequencies of the two alleles will remain the same from generation to generation. To prove it, finish the Hardy-Weinberg derivation worksheet. Assuming Hardy-Weinberg equilibrium is in effect we can calculate the allelic frequencies from the genotypic frequencies. To calculate the allelic frequency from the genotypic frequency, first figure out from the genotypes where all the A alleles are found, (AA and ½ of the alleles of Aa individuals will be A). So (where f stands for frequency), p = f(AA) + ½ f (Aa). Since p2 represents the frequency of AA and 2pq represents the frequency of Aa then when we substitute those frequencies into the equation we get, p = p2 + ½(2pq). Using analogous calculations we can calculate the frequency of the a (q)

Identify why the Hardy-Weinberg Equilibrium is difficult or impossible to demonstrate with living organisms. migration. white and brown/white speckled). Design and perform a test of a disruption to Hardy-Weinberg Equilibrium. but the Hardy-Weinberg Equilibrium says that if you remove mutation. selection. for instance. the allele frequencies will stay the same: you won’t see a change in the genotypes in the population. In absence of any factors that change the allelic frequencies. Students know the conditions for the Hardy-Weinberg equilibrium in a population and why these conditions are not likely to appear in nature. we are going to work not with real organisms but with a model—in this case dried beans. The Hardy-Weinberg equilibrium also assumes random mating in the population. Therefore. Materials • Opaque Tupperware containers or coffee cans with lids • Three different colors of beans that are approximately the same size (e.. 3.allele. For each group you will need 2. 4. completely isolated. students should be able to 1. for the purposes of our demonstration. If we could eliminate natural selection. given the frequency of phenotypes. 2. brown. Discuss the conditions under which a population is in Hardy-Weinberg Equilibrium.g. 5. and once again this is a situation that is rarely found in nature and difficult to achieve in the lab. Calculate genotypic and allelic frequencies. Over a long period of time. Derive the Hardy-Weinberg Equation. 7f. 840 speckled beans. Students know why alleles that are lethal in a homozygous individual may be carried in a heterozygote and thus maintained in a gene pool. do you think the population would change based solely on the recombination of genes that happens with sex? It’s almost impossible to test such a thing with living organisms. and chance effects from a mating population. Motivation What causes organisms to change from one generation to the next? We know that environmental pressures are a key force in natural selection.000 beans (180 white beans. Determine if a population is in Hardy-Weinberg Equilibrium. This lab uses beans rather than living organisms to simulate such a set up. would mating organisms change over time? What about mutation? Imagine a large population of breeding fruit flies. the genotypic and allelic frequencies will remain the same from generation to generation. and 980 red beans) Associated California Biology Standards 7b. . How right is HardyWeinberg? Objectives Upon completion of the lab. Students know how to solve the Hardy-Weinberg equation to predict the frequency of genotypes in a population. 6. 7e.

Do one color bean for all containers before moving to a second bean. According to the Hardy-Weinberg equation. 7. Hand out bean containers. 5. and distribute materials. and for the last counts of types #1 and #3. 4. Estimate this as 100 beans. Use the attached handout to help them along the way. Many of them are also lethal and many of them have also been present in the human gene pool for hundreds of thousands and possibly even millions of years. Test Preparation 1. Use this procedure to count out 180 beans of type #1. 6. weigh 100 beans and pull out 20 from that container. 840 of type #2. Extension Activity 1. and 980 of type #3 (according to the student handout). 2. You can count out the 40 beans for type #2 separately. Evaluation The following questions are listed under the Analysis section of the student handout and maybe used as part of a report. 3. Count 100 beans of one color by hand and weigh them. the dominant trait is represented by (A) p (B) q (C) q2 (D) p2 (E) 2pq . Place a cup on a scale and zero the scale. What conditions must be present for Hardy-Weinberg Equilibrium to be in effect? 2. Fill it with beans until it reaches the weight of the beans in step 1. Explain why evolution can not happen if the Hardy-Weinberg equilibrium is true in a population. or assessment 1. Many deleterious genes are recessive and therefore are expressed only in the homozygote. Distribute and explain copies of Hardy Weinberg simulation (see below). Use the following summary and test preparations sections to review the main ideas of the lesson. class discussion. depending on class size. How do you explain the persistence of these genes over such long periods of time in the face of such intense selective pressure against them? 3. Divide the students into groups of 5 or 6.Procedure 1. Have your students derive the Hardy-Weinberg Equation.

In a population of 1. What percent of the population has hybrid brown eyes? (A) 3% (B) 9% (C) 21% (D) 42% (E) 49% 3.000 people. the relative frequencies of the two alleles in the gene pool (E) a decrease in 2 pq below the value expected by the Hardy-Weinberg theorem . the most likely effect of assortative mating on the frequencies of alleles and genotypes for a locus would be (A) a decrease in p2 compared to q2 (B) a trend toward zero for q2 (C) convergence of p2 and q2 toward equal values (D) a change in p and q. 90 have blue eyes.2. In terms of the algebraic symbols used in the Hardy-Weinberg equation (p and q).

brown and speckled . Homozygous dominant individuals = (0. The first thing we have to do is to calculate the frequency p of the dominant allele A and the frequency q of the recessive allele a. Finally. and these represent individuals that are homozygous dominant for the trait (genotype AA).000 = Homozygous recessive individuals = (0. first of all determine the total number of alleles for a given gene in a population of 2000 diploid individuals. representing a population of 2. genotypes. Number of unique alleles = ___________ Total number of alleles = ______________ Next.000 dried beans. Done That: A Hardy-Weinberg Simulation Each of these containers contains approximately 2. so we have to know how many of these individuals we have. (This starting information is summarized in Table 1. how many of these are dominant (A) alleles? These alleles are found in homozygous dominants and in heterozygotes.Name:__________________ Student Handout—Bean There. Color Genotype Relative frequency % White aa 9 Speckled Aa 42 Brown AA 49 Procedure (A) Calculating allele frequencies p and q arithmetically. Bean colors.000 = .09) x 2. To calculate p by simple arithmetic. and genotype frequencies.and each color represents one of the three possible genotypes for a certain trait. forty-nine percent of the beans are brown. The beans are of three colors -white. and these represent individuals that are homozygous recessive for the trait (genotype aa). This can be done either by using simple arithmetic or by using the Hardy-Weinberg equation.49) x 2.000 diploid organisms. Forty-two percent of the beans are speckled and these represent individuals that are heterozygous for the trait (genotype Aa).) Table 1. Nine percent of the beans are white.42) x 2.000 = Heterozygous individuals = (0.

it is often impossible to distinguish between the phenotypes of homozygous dominants and heterozygotes.How many dominant (A) alleles are there in this population? Remember. Do we get the same answer for p and q by using simple arithmetic (Section A) as we get by using the Hardy-Weinberg equation (Section B)? Is p + q = 1 in both cases? For genes that show dominance. In these cases. the frequency of the recessive allele a in the population. by simple arithmetic. Shake your can vigorously to make sure your beans are thoroughly mixed. q =____________ You can check you answer by calculating q the same way we calculated p above. p. p = the frequency of AA genotypes + 1/2 the frequency of Aa genotypes In our populations. This is equivalent to (p+q)=1 (If you need to prove that to yourself. we might take the square root of the frequency of homozygous recessives to determine q. without looking. reach into the can and draw out the first two beans your fingers encounter. can be determined by dividing the total number of A alleles by the total number of alleles. Recall that the Hardy-Weinberg equations tells us that 1=p2+2pq+q2. Knowing that p+q =1. because they look the same. the frequency of A alleles in the population. p= Total number of A alleles = _____________ Total number of alleles Now repeat this procedure to get q. and the estimate the value of p by the equation p=1 – q. Then. We are now ready to begin our random mating simulation. Total number of A alleles = _____________ Total number of a alleles = _____________ Finally. p calculated in this way would be ______________. Calculate q using the value for p you calculate above. each homozygous dominant individual has 2 such alleles. q= Total number of a alleles = _____________ Total number of alleles (B) Calculating allele frequencies p and q from the Hardy-Weinberg equation We can calculate p using the frequencies of the genotypes from Table 1. square both sides of the p+q=1 equation and see what you get). Use a tally mark to . but each heterozygote has only 1. (C) The simulation. q = the frequency of aa genotypes + 1/2 the frequency of Aa genotypes In our populations. q calculated in this way would be ______________.

To do this. Random mating tallies. multiply that number by the fractional expected genotypic ratio for each genotype. Mating Pairs Genotypes Mating (color x color) Tallies brown x brown AA x AA brown x speckled AA x Aa brown x white AA x aa speckled x Aa x Aa speckled speckled x white Aa x aa white x white aa x aa (D) Using the results to calculate genotype frequencies. and enter these totals in Table 4. or 0) and record them in Table 3. Put the beans back in the can and draw out another pair. Next. use Punnett squares and your knowledge of Mendelian genetics to determine the genotypic ratios that we would expect to find in the offspring that would be produced by each of the six possible mating combinations in our population. 1/4. Total the number of offspring of each of the three possible genotypes. Table 2. The second one is filled in for you. Express these genotypic ratios as fractions (1. and multiply the number of times each mating type was drawn by four to get the total number of offspring produced by matings of that particular type. Determine the representation of each genotype among the offspring produced by these matings. Repeat this procedure until you have tallied a total of twenty-five matings on Table 2.record the colors of the beans you have withdrawn. and put them back into the can. on Table 2. 1/2. . Then fill out Table 4 with the fractions of each genotype and the number of times you drew each of the six possible mating types. Finally. assume that each mating produces four offspring. and the genotypes these colors represent. Tally the colors and genotypes of this pair. This will give you the number of offspring of a particular genotype that we can expect to be produced from matings of each of the six possible mating types.

Enter these genotype frequencies in Table 5. Do they correspond to the genotype frequencies with which we began. # = numbers of offspring of various genotypes. as recorded in Table 1? . the genotype frequency expressed as a percentage for each of the three possible genotypes will be numerically equal to the number of offspring for each of these genotypes. Therefore. Representation of various genotypes among offspring. the grand total of all offspring produced is equal to 100.Table 3: Predicted Outcomes From Potential Crosses AA x AA AA xAa AA x aa Aa x Aa AA Aa AA Aa Aa x aa aa x aa Genotypic ratios expressed as fractions: AA __ AA ½ AA___ Aa __ Aa ½ Aa___ aa __ aa 0 aa___ AA___ Aa___ aa___ AA___ Aa___ aa___ AA___ Aa___ aa___ Table 4. Mating Type Times Drawn Offspring per mating Total offspring Ratio AA AA X AA AA X Aa AA X aa Aa X Aa Aa X aa aa X aa 4 4 4 4 4 4 Ratio Aa Ratio aa # AA # Aa # aa Ratio = genotypic ratios of offspring expressed as fractions. Since you drew 25 mating pairs and each mating produced four offspring.

Do not add beans to a coffee can that were not in that can to begin with. Put any beans that you remove from a can back in the same coffee can they came from. Has either p or q changed in the new generation? Have we demonstrated successfully that genetic recombination as a result of sexual reproduction can not by itself result in any change in allele frequencies or genotype frequencies from one generation to the next? Remember that our demonstration was designed to exclude the effects of various factors that can alter allele frequencies. suggest some ways in which we might change our procedure so as to eliminate or at least reduce the impact of these effects on our results. and the frequency q of the recessive allele a in the same group. been entirely successful in excluding the influence of chance effects on allele and genotype frequencies? If you don't think that we have. What conditions must be present for Hardy-Weinberg Equilibrium to be in effect? 2. Suggest an experiment or exercise using our beans and coffee cans that would demonstrate the effect of migration on allele and genotype frequencies. How could we demonstrate the impact of selective pressures? Of non-random mating? (This matter of non-random mating is particularly intriguing. Many of them are also lethal and many of them have also been present . and if so how? What if you did the opposite and permitted only beans representing different genotypes to mate? Would this procedure alter genotype frequencies in the next generation. THIS IS VERY IMPORTANT!) Analysis Answer the following questions on a separate sheet of paper. Design and carry out at least one of these exercises and report your results to the class. (E) Demonstrating factors that skew Hardy-Weinberg predictions. Many deleterious genes are recessive and therefore are expressed only in the homozygote. How successfully has this been done? How would you be sure that statistically you had been successful? Have we. for example. and if so how? Would either procedure have any effect on allele frequencies?). What would happen if. 1. you prohibited matings between beans representing different genotypes and permitted only beans representing the same genotype to mate? Would this procedure alter genotype frequencies in the next generation. (Whatever you do. make sure that you leave the coffee cans as you found them.Table 5: Genotypic frequencies Genotype Number of Offspring Relative Frequency (%) AA Aa aa Value p______________ Value q ________________ Now calculate the frequency p of the dominant allele A among the offspring produced by our simulated random matings. You may do this either arithmetically or by using the Hardy-Weinberg equation. for example.

How do you explain the persistence of these genes over such long periods of time in the face of such intense selective pressure against them? 3. .in the human gene pool for hundreds of thousands and possibly even millions of years. Explain why evolution can not happen if the Hardy-Weinberg equilibrium is true in a population.

the possible combinations are Male pA p2 AA pq Aa Female Gametes pA qa qa pq Aa q2 aa This table shows the relationship between the allelic frequencies (p and q) and the genotypic frequencies (p2. the mating frequency of AA x AA. Each individual will carry two alleles of every gene. and E. the total of Column B should equal the totals of D. first consider all the possible matings from every genotypic outcome above. 2pq. and is free from evolutionary forces. the combined totals of C. so the frequency of that genotype in the next generation is p4. tally down to get the totals in each column. In a hypothetical population where the frequency of allele A is p and the frequency of allele a is q. D. C. mates randomly. To do this. and E combined. Therefore. which form the basis of the HWL. we can assume that all matings are random and therefore will occur with equal frequency. the frequency of the genotype AA is p2. since we are assuming that this population is subject to HWL. . for instance. For the above example. 2pq and q2. that would mean that after many generations the frequency of AA is still p2 and the frequency of Aa is still 2pq. The Hardy-Weinberg Law (HWL) states that when a population is in equilibrium. Columns C-E are the genotypic frequencies resulting in the next generation. Fill in the chart below. which should come out the same as the frequencies of the original generation.Hardy-Weinberg Law Derivation The Hardy-Weinberg Law: 1=p2+2pq+q2 Recall that alleles are alternative forms the same gene. Therefore in a population with just 2 alleles of a gene. If Hardy-Weinberg is correct. which makes up the entire population of the next generation. is p2 x p2 or p4. which is already filled in for Column B. AA x AA matings don’t occur more often than aa x aa matings. These matings are listed in Column A below. For example. 100% of the offspring will have the genotype AA. This means that the frequency of mating between any two genotypes is the product of how frequently these genotypes are present in the population. In our example of AA x AA. For instance. Afterwards. one from their mother and one from their father. the genotypic frequencies will be in the proportion p2. Next. Now that we know how frequent each of these matings are. and q2). We can demonstrate this by considering a hypothetical randomly mating population from table above. This composition of alleles for all individuals in a population is called the gene pool. the frequency of the genotype Aa is 2pq. The HWL states that the genotypic frequencies remain constant generation after generation if the population is large. we can determine the alleles that the offspring of that mating will contribute to the gene pool. Put another way. which is located at a specific position on a specific chromosome. each genotype passes on both alleles that it possesses with equal frequency. The genetic composition of each individual is that individual’s genotype. should still result in the same Hardy-Weinberg equation: 1=p2+2pq+q2.

(Recall that the chi-square test gives us the probability that the difference between what we observed and what we expected under the HWL is due to chance.995. obtain gene frequencies and calculate the expected genotypic frequencies and compare those frequencies with the observed frequencies of the genotypes using a chi.005 and p= 1-q = 0. which are usually rare and recessive.995 x 0. If q2 = 0.000025. This can illustrate how a rare allele is maintained in a population. An example: tyrosine-negative albinism is present in 1 in 40.000 individuals. This point is also illustrated by the distribution of genetic diseases in humans. or 0.005 = 0.00995 (almost 1% of the population).000025 then q= 0. This rare allele will be maintained not only in the homozygote but in the heterozygote population too. the homozygote for that allele is the rarest of the genotypes.square test.) . The heterozygote frequency is 2pq = 2 x 0. To determine if HWL is in effect in a population.A Type of mating Male x female p2 AA x p2 AA p2 AA x 2pq Aa 2pq Aa x p2 AA p2 AA x q2 aa q2 aa x p2 AA 2pq Aa x 2pq Aa B Mating frequency p4 C AA p4 D Offspring frequencies Aa - E aa - - 2pq Aa x q2 aa q2 aa x 2pq Aa q2 aa x q2 aa total - p2 AA +2pq Aa + q2 aa = 1 Implications of HWL When the frequency of one allele is low. The heterozygote carrier will be 1 out of every 100 people.

first consider all the possible matings from every genotypic outcome above. Each individual will carry two alleles of every gene. since we are assuming that this population is subject to HWL. the mating frequency of AA x AA. In our example of AA x AA. 2pq and q2. Now that we know how frequent each of these matings are. The genetic composition of each individual is that individual’s genotype. so the frequency of that genotype in the next generation is p4. is p2 x p2 or p4. C. each genotype passes on both alleles that it possesses with equal frequency. This means that the frequency of mating between any two genotypes is the product of how frequently these genotypes are present in the population. AA x AA matings don’t occur more often than aa x aa matings. we can determine the alleles that the offspring of that mating will contribute to the gene pool. mates randomly. If Hardy-Weinberg is correct. 100% of the offspring will have the genotype AA. D. and E. We can demonstrate this by considering a hypothetical randomly mating population from table above. tally down to get the totals in each column. the genotypic frequencies will be in the proportion p2. Therefore. and is free from evolutionary forces. Put another way. This composition of alleles for all individuals in a population is called the gene pool. the combined totals of C. In a hypothetical population where the frequency of allele A is p and the frequency of allele a is q. which is located at a specific position on a specific chromosome. which form the basis of the HWL.Hardy-Weinberg Law Derivation The Hardy-Weinberg Law: 1=p2+2pq+q2 Recall that alleles are alternative forms the same gene. For instance. and E combined. The Hardy-Weinberg Law (HWL) states that when a population is in equilibrium. we can assume that all matings are random and therefore will occur with equal frequency. the frequency of the genotype AA is p2. To do this. and q2). the frequency of the genotype Aa is 2pq. which is already filled in for Column B. These matings are listed in Column A below. . which should come out the same as the frequencies of the original generation. that would mean that after many generations the frequency of AA is still p2 and the frequency of Aa is still 2pq. Therefore in a population with just 2 alleles of a gene. should still result in the same Hardy-Weinberg equation: 1=p2+2pq+q2. Columns C-E are the genotypic frequencies resulting in the next generation. Fill in the chart below. For the above example. Next. For example. the total of Column B should equal the totals of D. The HWL states that the genotypic frequencies remain constant generation after generation if the population is large. the possible combinations are male pA p2 AA pq Aa female Gametes pA qa qa pq Aa q2 aa This table shows the relationship between the allelic frequencies (p and q) and the genotypic frequencies (p2. Afterwards. 2pq. which makes up the entire population of the next generation. for instance. one from their mother and one from their father.

The heterozygote carrier will be 1 out of every 100 people. This rare allele will be maintained not only in the homozygote but in the heterozygote population too. or 0.) .005 = 0. If q2 = 0.000 individuals.005 and p= 1-q = 0. the homozygote for that allele is the rarest of the genotypes.00995 (almost 1% of the population). which are usually rare and recessive.square test.000025. (Recall that the chi-square test gives us the probability that the difference between what we observed and what we expected under the HWL is due to chance. To determine if HWL is in effect in a population.000025 then q= 0.995 x 0. An example: tyrosine-negative albinism is present in 1 in 40. The heterozygote frequency is 2pq = 2 x 0. obtain gene frequencies and calculate the expected genotypic frequencies and compare those frequencies with the observed frequencies of the genotypes using a chi.995.A Type of mating Male x female p2 AA x p2 AA p2 AA x 2pq Aa 2pq Aa x p2 AA p2 AA x q2 aa q2 aa x p2 AA 2pq Aa x 2pq Aa B Mating frequency p4 4p3q 2p2q2 4 p2q2 4pq3 q4 (p2 +2pq + q2)2=1 2 2 C AA p4 2 p3q p2q2 p (p +2pq + q ) = p2 2 D Offspring frequencies Aa 2 p3 q 2p2q2 2 p2q2 2 pq3 2pq (p +2pq + q ) = 2pq 2 2 2 2 E aa p2q2 2 pq3 q4 q (p +2pq + q2) = q2 2pq Aa x q2 aa q2 aa x 2pq Aa q2 aa x q2 aa total p2 AA +2pq Aa + q2 aa = 1 Implications of HWL When the frequency of one allele is low. This can illustrate how a rare allele is maintained in a population. This point is also illustrated by the distribution of genetic diseases in humans.