1. What is the Makeup of the Community of Organisms Living on Rock Substrate Near the Post in the Long Beach High School Pond?
Matthew Amato, Joseph Carrasco, Carmen Sabedra & Cody Onufrock
Blast results in table and pictures of gracilaria tikvahiae
Abstract
The question posed is, What is the Makeup of the Community of Organisms Living on rock substrate near the post in the Long Beach High School Pond? Organisms were collected and identified by using DNA barcoding. With the DNA from the collected organisms, via gel electrophoresis, traces of viable DNA were discovered . When gels were run, only one sample of six had worked. Because of the wide range of organisms collected, it was concluded that the community surrounding the post is diverse and strong enough to support a multitude of different species. The most important finding from this research was coincidentally, the only organism that worked during gel electrophoresis.( This organism, PAZ-01, was the only organism that was not identifiable as an animal or plant using taxonomic keys)
Introduction
Organisms were collected within 6 inches of the designated post, which was a concrete pipe that allows water from the Reynolds Channel bay to drain in and out of the pond at the Long Beach High School.
Surrounding the pipe, are rock debris perhaps placed there during pipe construction. These rock debris are seemingly teeming with life.
There are many crevices in the rocks and they have been in the water for a number of years, which is why samples were taken from these specific rocks.
Rocks were taken from the pond bed, and analyzed the organisms and plant life living on them.
Reynolds Channel supports the life of many isopods, worms, plants such as Ulva lactuca, and Amphipods, especially of the Talitridae family. Materials & Methods
Tube Rack
Forceps
Alcohol Prep Wipes
Microcentrifuge Tubes
Microcentrifuge
Distilled Water
Micropipettes (sizes 10, 100, and 1000)
Results
By blasting the DNA sequence (pictured above abstract), it was discovered that the sample was a piece of the organism Gracilaria tikvahiae, which is commonly found in bay areas like the Long Beach High School Pond, which is full of water that drains in and out from Reynolds Channel. 15 samples were also collected from the pond these include: isopods, worms, plants such as Ulva lactuca, and Amphipods, especially of the Talitridae family.
Photo 1: DNA extracting process
Photo 2: Map and coordinates of where organism were collected
Discussion
A variety of species were found that were difficult to extract from the rock substrate, but the DNA that was successfully extracted and analyzed was from the organism Gracilaria tikvahiae, more commonly known as Graceful Red Weed. Gracilaria tikvahiae is known for appearing in nutrient rich bay areas, which help it to survive. The optimum growth of Gracilaria tikvahiae occurs between 24°C - 30°C. Graceful Red Weed produce carrageenan, which is commonly used in the food industry for gelling, thickening, and stabilizing properties. Nutrient loading of the bay is likely increasing the population of the organism. DNA was extracted from other organisms as well but amplification was not sufficient
to show substantial amounts of DNA on the gels.
Acknowledgements
We would like to make a personal thank you to our instructor Cody Onufrock and the Cold Spring Harbor DNA Learning Center.
Lysis solution was added and used mortar used to break open cell wall and release cell contents into liquid. Silica resin was then added to a new centrifuge tube to bind DNA, and removed cell parts from our DNA. DNA was amplified by PCR, allowing for the copying of selected gene fragment millions of times in order to have a quantity that is able to be sequenced. This process allowed the checking of samples for the presence of viable DNA to see if the Barcoding Process was successful.
Insect COI Primer
Agarose Gel
Sybr Green Dye
Wash Buffer
PCR Product
Photo 3: sample Z
Photo 4: sample J1
Photo 5: sample B1
Photo 6: sample E1

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