# Alligator Snapping Turtle eDNA Analysis from Filtered Water Samples Canonical: https://abierto.us/opportunities/1132837 - Solicitation number: 1132837 - Notice type: Sources sought - Status: Closed. Deadline was August 6, 2024 at 11:59 PM EDT - Department: Department of Agriculture - Agency: Forest Service - Contracting office: Usda-Fs, Csa East 8 (124447) - NAICS: 541380 Testing Laboratories and Services - Product or service code: B504 Chemical/Biological Studies and Analysis - Set-aside: Total small business set-aside - Place of performance: Texas - County: Walker County (FIPS 48471). https://abierto.us/counties/walker-county-tx-48471 - First posted: July 26, 2024 - Last posted: July 26, 2024 - SAM.gov: https://sam.gov/workspace/contract/opp/f44eb033ad0b4ca5a4d097e46b6c336b/view ## Description The proposed work is to analyze samples for a single species, the Alligator Snapping Turtle, for environmental DNA (eDNA). Water samples (135 total) were collected from lakes and streams and filtered in the laboratory using analytical filter funnels containing 0.45um filters. Filters, containing eDNA, were removed from filter funnels and placed in labeled microcentrifuge tubes and the tubes were placed in a deep freeze. Each microcentrifuge tube contains 2 filters from the same water sample. Samples will be shipped in an insulated container, containing cold packs, to a laboratory for analysis. Each tube should be considered as an independent sample. Laboratory is required to detect sequences of Alligator Snapping Turtle DNA from filter medium and ensure no cross contamination of samples. Laboratory must provide a detailed report showing CT scores for each run (3 runs per sample) and if each sample has a positive or negative result for Alligator Snapping Turtle DNA. All sample results must remain distinct for future statistical analysis. The contractor must extract water filters using the Zymo Quick Miniprep Kit extraction kits (Zymo Research, Orange California) following a modification of the Spear et al. (2015) protocol. Next, they must amplify the DNA in a qPCR reaction using primers and thermocycler profile modified from Feist et al. (2018) to work with samples throughout the range of Alligator Snapping Turtles. The contractor must visualize amplification using a QuantStudio3 machine (Thermo Fisher Scientific, Waltham MA). The samples must be run with a negative water control and internal positive control designed to capture the genetic variation present across the range of Alligator Snapping Turtles (interspecific and intraspecific) Vendors/laboratories must submit detailed capability statements that demonstrate how the same testing methods described above could be used. Capability statements must also show how results would ensure consistency with prior testing in order to validate the overall research study findings. General capability statements should not be submitted. Vendors must demonstrate that they can perform the requested analysis following the requested methodology. Submit detailed capability statements by e-mail to Laniece Nixon at laniece.nixon@usda.gov with a courtesy copy to Keith Friot at keith.d.friot@usda.gov ## Publications - July 26, 2024: Sources sought, due August 6, 2024 at 11:59 PM EDT. Notice f44eb033ad0b4ca5a4d097e46b6c336b. https://sam.gov/workspace/contract/opp/f44eb033ad0b4ca5a4d097e46b6c336b/view ## Points of contact - Laniece Nixon, laniece.nixon@usda.gov - Keith Friot, Keith.d.friot@usda.gov, 9104751968 --- Source: SAM.gov Contract Opportunities bulk extract. Confirm deadlines on SAM.gov before responding. Cite https://abierto.us/opportunities/1132837.