General Guidelines:

The easiest way to do the poster is to break it up into pieces and everybody in the group take one or two pieces. Group leaders are responsible for assigning tasks and coordinating the work of putting the poster together. They are not responsible for doing all the work themselves! Produce a poster that you will be happy to have displayed in the hall.

Please remember that everybody in the group is responsible for all the information on the poster. Be sure you all understand what was done.

WEDNESDAY LAB
 

THURSDAY LAB
 

FRIDAY LAB
 

Posters are similar to journal articles, but in a condensed way. All the information has to be easy to understand and presented in short segments on the poster board. This is the most significant difference between a poster and a journal article. You can use headings with a few main points explained, short paragraphs, or other ways to break up the text. In the Methods, you may want to use an outline or a table. The font should be large and easy to read (no script!). The main parts of a poster are:
 

1. Title:  includes name of project, people who did the work (your group) and location. Feel free to change the title I used above.
 

2. Introduction:
Explain why the project was performed (similar to a "statement of purpose" when writing a paper). Like a paper, you need to give background information so people can understand the results. We had one main goal, identification of unknown bacterial samples by using DNA sequencing. But we had a reason for choosing a certain segment of DNA.

PCR of 16s rDNA
Posters need to explain the basics about the PCR technique and how it can be used to amplify one small segment of a genome. In our case, it was a portion of the bacterial 16s rDNA gene. What is the significance of that segment? How do researchers use sequence information to identify microorganisms and to establish phylogenetic relationships? It will be important to give enough background so people reading the poster in the hall will understand the use of ribosomal DNA in identification.

Sequencing
Molecular lab has done PCR before but this is the first time we have had the amplified product sequenced. Explain the process of "cycle sequencing". Dana gave me a copy of her powerpoint disk so you can borrow that to look at again. A picture or drawing might be useful here. How does a capillary sequencer work? What type of information is obtained?

Identification
How did we identify our unknown bacteria? Explain the things we looked for when examining our chromatograms, such as N bases or two bases assigned to one peak. Describe how we determined the identity of our unknowns. What was the purpose of using the BLAST program at NCBI? What information did we get from the BLAST search? What are the pros and cons of using public databases?

As a caution, I urge you to not get all your information from the internet. Make sure your information can be referenced to a text or journal article or other reputable scientific source.

Remember that figures are a good way to explain things without having big blocks of text. So you might consider incorporating figures, charts, diagrams, etc. into the Introduction. Some figures might serve double duty in other sections such as Methods.
 

3. Methods:
To recap, our Methods for cloning the rDNA PCR segment included:

isolation of bacterial DNA, PCR, verification of PCR product (gel), purification of PCR product from agarose gel (Novagen protocol), verification of purification (gel), and turning samples over to the Grice sequencing facility. They did the "cycle sequencing" reaction, which is PCR using the dideoxy nucleotides. Samples were run on a capillary sequencer.

• The experiment starts with PCR of a certain region of the 16s rDNA gene. You have the original paper as well as a handout giving the changes we made to that protocol. You also have the Novagen protocol for isolating DNA from a preparative agarose gel.
• Then the DNA was handed over to the sequencing facility at Grice. A proper way to designate this would be : Core Sequencing Facility, Grice Marine Lab (College of Charleston, Charleston SC). Our posters should include specifics on the kits used for cycle sequencing (and refer to any useful figures from the Introduction) and the type of capillary sequencer (CEQ 8000 from Beckman Coulter).

4. Results:
A figure should have a Figure Number, possibly a title,  and a Figure Legend. The Figure Legend will tell what the lanes on a gel represent, what the axes on the graphs are, etc. Tables should have a number and a title. I suggest you look around the building at posters to see how data is presented. All posters need a figure or table that summarizes the identification results. It should include the unkniwn code # and identification to at least the genus level.
What happens if I can't identify my bacterium to genus level? Well, you probably can't choose between two genera. Therefore, find a traditional microbiological test that can distinguish between the two possibilities. You can confer with the people who have had/are taking microbiology but you should have an idea before you take up their time. This information should be provided on the poster, but it can be in a separate section  if you want.

•  Include gel photos large enough to see the important points. The gel pictures should be properly labeled.
• The following gel photos are available: verification of PCR product, preparatory gel for isolating the DNA fragment, verification of the purification procedure.
• A sequence printout (chromatogram) can be made using an inkjet printer.

5. Discussion or Conclusions: Discuss your experimental data.

• In general, how successful was our approach for identifying unknown bacteria?
• We would like to be able to identify unknown bacteria based on sequence information. What are the traditional methods used for bacterial identification? What are the advantages and disadvantages of each approach? Someone with microbiology experience may be best able to address this.
• Are there ways that we might improve our chances of obtaining the correct identification? What are they and how would they help?

This is the information I would like on everyone's poster. But please feel free to elaborate on topics that interest you! I am looking forward to learning more from your posters. So have some fun with this assignment, and take advantage of the opportunity to expand your horizons.
 

7. Poster Presentation:

Each group will present its poster in class, and the other group will be the audience. You will present the section(s) of the poster that you worked on. Please practice your presentation so you will be confident in what you say. Ideally, everyone should practice with the group so you can coordinate what each person will say. This is important, because sometimes Methods material is discussed in Intro, or Results are discussed in Methods. That's OK, as long as you leave your colleagues with something to talk about! So work on this ahead of time and practice your part so it flows smoothly. Your presentation will be a reflection of how well you understand your section and the project as a whole, so show your audience that you know what's going on!

Public speaking is an important skill for a scientist to have, but it makes many people nervous. Practice is the best remedy. So practice by yourself and with the group. Don't be nervous when you give your part because you will be speaking before a very friendly and understanding audience.