Biological Oceanography Group (BOG) Protocol Collection

Member of Marine Biodiversity Observation Network (MBON) - CeNCOOS MBON

Website describing protocols: https://mbari-bog.github.io/MBON-Protocols/

BeBOP formatted detailed protocols with DOIs below:

Authors

• Jacoby Baker
• Kobun Truelove
• Katie Pitz
• Francisco Chavez

Protocols Currently in Use

Sample Collection Protocols

• eDNA sampling using a peristaltic pump with preservation in DNA/RNA Shield solution

Extraction Protocols

• Environmental DNA (eDNA) extraction using Qiagen DNeasy 96 Blood and Tissue Kit V.3

PCR Protocols

• Environmental DNA (eDNA) 18S metabarcoding PCR Protocol V.3
• Environmental DNA (eDNA) COI metabarcoding PCR Protocol V.2
• Environmental DNA (eDNA) 12S Metabarcoding PCR Protocol (with Platinum SuperFi II Taq) V.3

Secondary PCR, Size selection, Library Prep and Sequencing Protocols

• Michigan State University (MSU) Illumina MiSeq sequencing with size selection protocol

Protocols Tested

PCR Protocols

Previously Used Protocols

Sample Collection Protocols

Extraction Protocols

PCR Protocols

• Environmental DNA (eDNA) 12S Metabarcoding PCR Protocol (Touchdown) V.2
• Environmental DNA (eDNA) 12S Metabarcoding PCR Protocol (Classic) V.1

Library Prep and Sequencing Protocols