--- author: Sam White toc-title: Contents toc-depth: 5 toc-location: left date: 2015-11-04 05:50:30+00:00 layout: post slug: dna-isolations-oly-fidalgo-2sn-ctenidia title: DNA Isolations – Oly Fidalgo 2SN Ctenidia categories: - 2015 - Olympia oyster reciprocal transplant tags: - DNA Isolation - E.Z.N.A. Mollusc Kit - Fidalgo 2SN - gDNA - olympia oyster - Ostrea lurida --- Isolated DNA from 24 2SN ctenidia samples from [Friday's sampling](https://robertslab.github.io/sams-notebook/posts/2015/2015-10-31-oyster-sampling-oly-fidalgo-2sn-2hl-2nf-reciprocal-transplants-final-samplings/) (#32 - 55). Samples were thawed at RT. 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: * Samples were incubated @ 60C for only 1hr, per Steven's recommendation (an attempt to prevent degradation) * No optional steps were performed * Used 300μL of MBL Buffer for all samples (this was more than the recovered volume of aqueous phase from each sample) * Single elution of 50μL Samples were stored @ -20C in: Oly gDNA Oly Reciprocal Transplant Final Sampling Box #1. Some notes: * Total time (including 1hr incubation): 4.5hrs. * Short incubation time did not completely digest samples * Partial tissue digestions led to difficulties in recovering entire aqueous phase, post chloroform treatment