Let me tell you about something.

Distractions and amusements, with a sandwich and coffee.

Trance opera
• Spente le Stelle

The Points of Significance column was launched in September 2013 in an effort to provide an educational resource to authors and provide practical suggestions about best practices in statistical analysis and reporting.

This month we launch a new column "Points of Significance" devoted to statistics, a topic of profound importance for biological research, but one that often doesnâ€™t receive the attention it deserves.

The "aura of exactitude" that often surrounds statistics is one of the main notions that the Points of Significance column will attempt to dispel, while providing useful pointers on using and evaluating statistical measures.

—Dan Evanko, Let's Give Statistics the Attention it Deserves in Biological Research

The column is co-authored with Naomi Altman (Pennsylvania State University). Paul Blainey (Broad) is contributing co-author.

Krzywinski, M., Altman, N. & Blainey, P. (2014) Points of Significance: Two factor designs *Nature Methods* **11**:1187-1188.

Krzywinski, M., Altman, N. & Blainey, P. (2014) Points of Significance: Nested designs *Nature Methods* **11**:977-978.

Blainey, P., Krzywinski, M. & Altman, N. (2014) Points of Significance: Replication *Nature Methods* **11**:879-880.

Krzywinski, M. & Altman, N. (2014) Points of Significance: Analysis of variance (ANOVA) and blocking *Nature Methods* **11**:699-700.

Krzywinski, M. & Altman, N. (2014) Points of Significance: Designing comparative experiments *Nature Methods* **11**:597-598.

Krzywinski, M. & Altman, N. (2014) Points of Significance: Non parametric tests *Nature Methods* **11**:467-468.

Krzywinski, M. & Altman, N. (2014) Points of Significance: Comparing samples—Part II — Multiple Testing *Nature Methods* **11**:355-356.

Krzywinski, M. & Altman, N. (2014) Points of Significance: Comparing samples—Part I — *t*-tests *Nature Methods* **11**:215-216.

Krzywinski, M. & Altman, N. (2014) Points of Significance: Visualizing samples with box plots *Nature Methods* **11**:119-120.

Krzywinski, M. & Altman, N. (2013) Points of Significance: Power and sample size *Nature Methods* **10**:1139-1140.

Krzywinski, M. & Altman, N. (2013) Points of Significance: Significance, *P* values and *t*-tests *Nature Methods* **10**:1041-1042.

Krzywinski, M. & Altman, N. (2013) Points of Significance: Importance of being uncertain *Nature Methods* **10**:809-810.

We've previously written about how to analyze the impact of one variable in our ANOVA column. Complex biological systems are rarely so obliging—multiple experimental factors interact and producing effects.

ANOVA is a natural way to analyze multiple factors. It can incorporate the possibility that the factors interact—the effect of one factor depends on the level of another factor. For example, the potency of a drug may depend on the subject's diet.

We can increase the power of the analysis by allowing for interaction, as well as by blocking.

Krzywinski, M., Altman, (2014) Points of Significance: Two Factor Designs *Nature Methods* **11**:1187-1188.

Blainey, P., Krzywinski, M. & Altman, N. (2014) Points of Significance: Replication *Nature Methods* **11**:879-880.

Krzywinski, M. & Altman, N. (2014) Points of Significance: Analysis of variance (ANOVA) and blocking *Nature Methods* **11**:699-700.

Krzywinski, M. & Altman, N. (2014) Points of Significance: Designing Comparative Experiments *Nature Methods* **11**:597-598.

Sources of noise in experiments can be mitigated and assessed by nested designs. This kind of experimental design naturally models replication, which was the topic of last month's column.

Nested designs are appropriate when we want to use the data derived from experimental subjects to make general statements about populations. In this case, the subjects are *random* factors in the experiment, in contrast to *fixed* factors, such as we've seen previously.

In ANOVA analysis, random factors provide information about the amount of noise contributed by each factor. This is different from inferences made about fixed factors, which typically deal with a change in mean. Using the F-test, we can determine whether each layer of replication (e.g. animal, tissue, cell) contributes additional variation to the overall measurement.

Krzywinski, M., Altman, N. & Blainey, P. (2014) Points of Significance: Nested designs *Nature Methods* **11**:977-978.

Blainey, P., Krzywinski, M. & Altman, N. (2014) Points of Significance: Replication *Nature Methods* **11**:879-880.

Krzywinski, M. & Altman, N. (2014) Points of Significance: Analysis of variance (ANOVA) and blocking *Nature Methods* **11**:699-700.

Krzywinski, M. & Altman, N. (2014) Points of Significance: Designing Comparative Experiments *Nature Methods* **11**:597-598.

It's fitting that the column published just before Labor day weekend is all about how to best allocate labor.

Replication is used to decrease the impact of variability from parts of the experiment that contribute noise. For example, we might measure data from more than one mouse to attempt to generalize over all mice.

It's important to distinguish technical replicates, which attempt to capture the noise in our measuring apparatus, from biological replicates, which capture biological variation. The former give us no information about biological variation and cannot be used to directly make biological inferences. To do so is to commit *pseudoreplication*. Technical replicates are useful to reduce the noise so that we have a better chance to detect a biologically meaningful signal.

*Nature Methods* **11**:879-880.

*Nature Methods* **11**:699-700.

Krzywinski, M. & Altman, N. (2014) Points of Significance: Designing Comparative Experiments *Nature Methods* **11**:597-598.

I was commissioned by Scientific American to create an information graphic that showed how our genomes are more similar to those of the chimp and bonobo than to the gorilla.

I had about 5 x 5 inches of print space to work with. For 4 genomes? No problem. Bring out the Hilbert curve!

To accompany the piece, I will be posting to the Scientific American blog about the process of creating the figure. And to emphasize that the *genome is not a blueprint*!

As part of this project, I created some Hilbert curve art pieces. And while exploring, found thousands of Hilbertonians!

Celebrate Pi Approximation Day (July 22nd) with the art of arm waving. This year I take the first 10,000 most accurate approximations (*m*/*n*, *m*=1..10,000) and look at their accuracy.

I turned to the spiral again after applying it to stack stacked ring plots of frequency distributions in Pi for the 2014 Pi Day.