--- author: Sam White toc-title: Contents toc-depth: 5 toc-location: left date: 2015-10-22 05:14:09+00:00 layout: post slug: dna-isolations-fidalgo-2sn-reciprocal-transplants-final-samplings title: DNA Isolations - Fidalgo 2SN Reciprocal Transplants Final Samplings categories: - 2015 - Olympia oyster reciprocal transplant tags: - ctenidia - DNA Isolation - E.Z.N.A. Mollusc Kit - Fidalgo 2SN - olympia oyster - Ostrea lurida - RNAlater --- The remaining Olympia oysters from Jake Heare's reciprocal transplant experiment have been retrieved from field sites and are awaiting sampling. The oysters have been stored in the cold room (temp?) for 6 days so far. Sampling scheme is as follows: 1. Assign unique number to oysters 2. Photograph with ruler for future shell measurements 3. Weigh oysters 4. Dissect ctenidia for DNA isolation 5. Dissect & discard viscera (e.g. digestive gland and gonad) 6. Weigh remaining body 7. Preserve remaining body in RNAlater 8. Weigh empty shells [ Mrunmayee photographed & initiated dissections of oysters #3 - 8](https://onsnetwork.org/mrunmayee/2015/10/21/opening-oysters/). I took over for oyster #9 -14. All oyster data is here (Google Sheet): [Oly reciprocal final sampling](https://docs.google.com/spreadsheets/d/1KjsfrNdUuXegcPx6UjoGfR7vM---LwpaNi6Nra3HMkI/edit?usp=sharing) DNA was isolated using the E.Z.N.A. Mollusc Kit (Omega Biotek) according to [the manufacturer's protocol](https://github.com/sr320/LabDocs/blob/master/protocols/Commercial_Protocols/Omega_Mollusc-DNA-Kit-Combo-May-2013-D3373.pdf) with the following changes: * No optional steps were performed * Ctenidia tissue was lysed for 3hrs @ 60C * Single elution of 50μL Samples were stored @ -20C in: Oly gDNA Oly Reciprocal Transplant Final Sampling Box #1.