Mio-Pliocene Plant DNA Barcoding

Regions: Inuvialuit Settlement Region

Tags: physical sciences, vegetation, fossils, paleontology, ancient plants

Principal Investigator: Spencer, Lee A (1)
Licence Number: 14525
Organization: Southern Adventist University
Licenced Year(s): 2009
Issued: Jun 03, 2009
Project Team: Art Chadwick, Ph.D. (Sedimentologist, Southwestern Adventist University), Karen Jensen, Ph.D. (Paleobotanist, Weimar College), Steve Austin, Ph.D. (Coal Petrologist, Austin Research Consulting, Inc.), Lucinda Hill, MD (Physician, Collection Assistant, Southern Adventist University), Leslie Schwarzer (Photographer, Camp Cook, Southern Adventist University), Paul Claus (Pilot, Ultima Thule Lodge), Donna Claus (Pilot's Wife, Ultima Thule Lodge), Lee Spencer, Ph.D. (Paleontologist, Principle Investigator, Southern Adventist University)

Objective(s): Collect plant fossils from the Miocene Ballast Brook and Pliocene Beaufort Formations (Ballast Brook River) and process for DNA.

Project Description: This licence has being issued for the scientific research application No.1072.

The researchers will collect plant fossils from the Miocene Ballast Brook and Pliocene Beaufort Formations (Ballast Brook River), Banks Island. They will transport the specimens to Southern Adventist University, Collegedale, Tennessee, USA, for DNA processing and analysis. A final report will be submitted to Aurora Research Institute detailing the processing and results.

Researchers will make camp on the northwestern end of Banks Island along the Ballast Brook River and plan to spend approximately four days studying the outcrops and collecting the best preserved specimens. In addition to individual specimens, one bulk frozen sample of about 10 liters (2.5 gal) will be collected and placed in a special anoxic chamber for transport back to the university labs. As the bulk samples thaw during transport, they will never be exposed to oxygen; nor will they be exposed to any contaminants including modern pollen. The individual specimens collected out of outcrop will be stored in a portable 12 volt freezer. Upon arrival at the university, the specimens will be transferred to the -70 C freezer where they will remain until processed.

DNA Extraction and Amplification:
All procedures for DNA extraction and PCR will be carried out in dedicated aDNA (ancient DNA) laboratory space (Mifflin 2003). Nucleic acids will be purified using a standard organic extraction procedure (using successive phenol, phenol/chloroform, and chloroform solutions). The nucleic acids will be either ethanol precipitated and re-suspended or concentrated in a Centricon concentrator (Millipore) prior to PCR. PCR reactions will be conducted with primer sets bracketing four plastid gene regions (rpoC1, rpoB, psbA-trnH and matK; with standard reaction conditions). PCR products will be separated by standard gel electrophoresis (1% agarose in TAE) and initially analyzed with a UVP EC3 documentation system equipped with VisionWorks Image Acquisition & Analysis Software. Following image capture and analysis of the PCR products, amplified DNA bands will be excised, extracted from agarose, and outsourced for DNA sequence determination.

Copies of all publications resulting from this study will be forwarded to the Aurora Institute. Additionally, there will be a preliminary report Spring, 2010, that will be submitted to the Institute.

The fieldwork for this study will be conducted on the northwestern end of Banks Island along the Ballast Brook River from July 10 to July 19, 2009.