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Before sequencing the DNA, we will do a PCR with primers that fit just before and just after the 16S DNA, in well conserved regions expected to be present in all bacteria.

The PCR replicates only the DNA region between our two selected primers a gazillion times so that only those regions will actually get picked up by the sequencing step in practice.

Eukaryotes also have an analogous ribosome part, the 18S region, but the PCR primers are selected for targets around the 16S region which are only present in prokaryotes.

This way, we amplify only the 16S region of bacteria, excluding other parts of bacterial genome, and excluding eukaryotes entirely.

Despite coding such a fundamental piece of RNA, there is still surprisingly variability in the 16S region across different bacteria, and it is those differences will allow us to identify which bacteria are present in the river.

The variability exists because certain base pairs are not fundamental for the function of the 16S region. This variability happens mostly on RNA loops as opposed to stems, i.e. parts of the RNA that don't base pair with other RNA in the RNA secondary structure as shown at: Code 1. "RNA stem-loop structure".

                A-U
               /   \
A-U-C-G-A-U-C-G     C
| | | | | | | |     |
U-A-G-C-U-A-G-C     G
               \   /
                U-A
|             ||    |
+-------------++----+
    stem        loop

Code 1.

RNA stem-loop structure

This is how the 16S RNA secondary structure looks like in its full glory: Figure 5. "16S RNA secondary structure".

height=800 — Figure 5.
16S RNA secondary structure
. Source.

Since loops don't base pair, they are less crucial in the determination of the secondary structure of the RNA.

The variability is such that it is possible to identify individual species apart if full sequences are known with certainty.

With the experimental limitations of experiment however, we would only be able to obtain family or genus level breakdowns.

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1881–1882 census of the Ottoman Empire by

Wikipedia Bot 0  1970-01-01

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 Pinned article: ourbigbook/introduction-to-the-ourbigbook-project

Welcome to the OurBigBook Project! Our goal is to create the perfect publishing platform for STEM subjects, and get university-level students to write the best free STEM tutorials ever.

Everyone is welcome to create an account and play with the site: ourbigbook.com/go/register. We belive that students themselves can write amazing tutorials, but teachers are welcome too. You can write about anything you want, it doesn't have to be STEM or even educational. Silly test content is very welcome and you won't be penalized in any way. Just keep it legal!

Video 1.

Intro to OurBigBook

. Source.

We have two killer features:

topics: topics group articles by different users with the same title, e.g. here is the topic for the "Fundamental Theorem of Calculus" ourbigbook.com/go/topic/fundamental-theorem-of-calculus
Articles of different users are sorted by upvote within each article page. This feature is a bit like:
- a Wikipedia where each user can have their own version of each article
- a Q&A website like Stack Overflow, where multiple people can give their views on a given topic, and the best ones are sorted by upvote. Except you don't need to wait for someone to ask first, and any topic goes, no matter how narrow or broad
This feature makes it possible for readers to find better explanations of any topic created by other writers. And it allows writers to create an explanation in a place that readers might actually find it.
Figure 1.
Screenshot of the "Derivative" topic page
. View it live at: ourbigbook.com/go/topic/derivative
Video 2.
OurBigBook Web topics demo
. Source.
local editing: you can store all your personal knowledge base content locally in a plaintext markup format that can be edited locally and published either:
- to OurBigBook.com to get awesome multi-user features like topics and likes
- as HTML files to a static website, which you can host yourself for free on many external providers like GitHub Pages, and remain in full control
This way you can be sure that even if OurBigBook.com were to go down one day (which we have no plans to do as it is quite cheap to host!), your content will still be perfectly readable as a static site.
Figure 2.
You can publish local OurBigBook lightweight markup files to either OurBigBook.com or as a static website
.
Figure 3.
Visual Studio Code extension installation
.
Figure 4.
Visual Studio Code extension tree navigation
.
Figure 5.
Web editor
. You can also edit articles on the Web editor without installing anything locally.
Video 3.
Edit locally and publish demo
. Source. This shows editing OurBigBook Markup and publishing it using the Visual Studio Code extension.
Video 4.
OurBigBook Visual Studio Code extension editing and navigation demo
. Source.
Internal cross file references done right:
Infinitely deep tables of contents:
Figure 6.
Dynamic article tree with infinitely deep table of contents
.
Live URL: ourbigbook.com/cirosantilli/chordate
Descendant pages can also show up as toplevel e.g.: ourbigbook.com/cirosantilli/chordate-subclade

All our software is open source and hosted at: github.com/ourbigbook/ourbigbook

Further documentation can be found at: docs.ourbigbook.com

Feel free to reach our to us for any help or suggestions: docs.ourbigbook.com/#contact

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