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A fairly common criticism of the hypothesis-testing approach to statistics is that the null hypothesis will always be false, if you have a big enough sample size. In the chicken-feet example, critics would argue that if you had an infinite sample size, it is impossible that male chickens would have *exactly* the same average foot size as female chickens. Therefore, since you know before doing the experiment that the null hypothesis is false, there's no point in testing it.

This criticism only applies to two-tailed tests, where the null hypothesis is "Things are exactly the same" and the alternative is "Things are different." Presumably these critics think it would be okay to do a one-tailed test with a null hypothesis like "Foot length of male chickens is the same as, or less than, that of females," because the null hypothesis that male chickens have smaller feet than females could be true. So if you're worried about this issue, you could think of a two-tailed test, where the null hypothesis is that things are the same, as shorthand for doing two one-tailed tests. A significant rejection of the null hypothesis in a two-tailed test would then be the equivalent of rejecting one of the two one-tailed null hypotheses.

For our problem, we need to create a test that will determine whether or not Brand A paper towel will absorb more water than Brand B paper towel. Our experiment consisted of measuring water, folding a paper towel, dipping it into water, noting how much water it had absorbed and then repeating the process with the other brand of paper towel.

involves the careful construction of two statements: the null hypothesis and the alternative hypothesis. These hypotheses can look very similar, but are actually different.

In a mathematical formulation of the null hypothesis there will typically be an equal sign. This hypothesis is denoted by *H*_{0}.

Does a probability of 0.030 mean that you should reject the null hypothesis, and conclude that chocolate really caused a change in the sex ratio? The convention in most biological research is to use a significance level of 0.05. This means that if the *P* value is less than 0.05, you reject the null hypothesis; if *P* is greater than or equal to 0.05, you don't reject the null hypothesis. There is nothing mathematically magic about 0.05, it was chosen rather arbitrarily during the early days of statistics; people could have agreed upon 0.04, or 0.025, or 0.071 as the conventional significance level.

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Another way your data can fool you is when you don't reject the null hypothesis, even though it's not true. If the true proportion of female chicks is 51%, the null hypothesis of a 50% proportion is not true, but you're unlikely to get a significant difference from the null hypothesis unless you have a huge sample size. Failing to reject the null hypothesis, even though it's not true, is a "false negative" or "Type II error." This is why we never say that our data shows the null hypothesis to be true; all we can say is that we haven't rejected the null hypothesis.

The significance level (also known as the "critical value" or "alpha") you should use depends on the costs of different kinds of errors. With a significance level of 0.05, you have a 5% chance of rejecting the null hypothesis, even if it is true. If you try 100 different treatments on your chickens, and none of them really change the sex ratio, 5% of your experiments will give you data that are significantly different from a 1:1 sex ratio, just by chance. In other words, 5% of your experiments will give you a false positive. If you use a higher significance level than the conventional 0.05, such as 0.10, you will increase your chance of a false positive to 0.10 (therefore increasing your chance of an embarrassingly wrong conclusion), but you will also decrease your chance of a false negative (increasing your chance of detecting a subtle effect). If you use a lower significance level than the conventional 0.05, such as 0.01, you decrease your chance of an embarrassing false positive, but you also make it less likely that you'll detect a real deviation from the null hypothesis if there is one.

When you reject a null hypothesis, there's a chance that you're making a mistake. The null hypothesis might really be true, and it may be that your experimental results deviate from the null hypothesis purely as a result of chance. In a sample of 48 chickens, it's possible to get 17 male chickens purely by chance; it's even possible (although extremely unlikely) to get 0 male and 48 female chickens purely by chance, even though the true proportion is 50% males. This is why we never say we "prove" something in science; there's always a chance, however miniscule, that our data are fooling us and deviate from the null hypothesis purely due to chance. When your data fool you into rejecting the null hypothesis even though it's true, it's called a "false positive," or a "Type I error." So another way of defining the *P* value is the probability of getting a false positive like the one you've observed, *if* the null hypothesis is true.

The relative costs of false positives and false negatives, and thus the best *P* value to use, will be different for different experiments. If you are screening a bunch of potential sex-ratio-changing treatments and get a false positive, it wouldn't be a big deal; you'd just run a few more tests on that treatment until you were convinced the initial result was a false positive. The cost of a false negative, however, would be that you would miss out on a tremendously valuable discovery. You might therefore set your significance value to 0.10 or more for your initial tests. On the other hand, once your sex-ratio-changing treatment is undergoing final trials before being sold to farmers, a false positive could be very expensive; you'd want to be very confident that it really worked. Otherwise, if you sell the chicken farmers a sex-ratio treatment that turns out to not really work (it was a false positive), they'll sue the pants off of you. Therefore, you might want to set your significance level to 0.01, or even lower, for your final tests.

After you do a statistical test, you are either going to reject or accept the null hypothesis. Rejecting the null hypothesis means that you conclude that the null hypothesis is not true; in our chicken sex example, you would conclude that the true proportion of male chicks, if you gave chocolate to an infinite number of chicken mothers, would be less than 50%.

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