Isolated gDNA from Ostrea lurida (Olympia oyster) adductor muscle & mantle samples collected by Brent & Steven on 20150812 using the E.Z.N.A Mollusc DNA Kit (Omega Bio-Tek) according to the manufacturer’s protocol, with the following adjustments:
- 29.8mg of adductor muscle
- 28.7mg of mantle
- Tissues homogenized in 350μL of ML1 Buffer with disposable mortar/pestle tubes using only three pestles strokes
- Homogenized tissue incubated in ML1 Buffer + Proteinase K @ 60C for 2.5hrs
- Added 310μL of MBL Buffer to adductor muscle sample and 265μL of MBL Buffer to mantle sample
- Added 620μL of 100% EtOH and 530μL of 100% EtOH to the adductor muscle and mantle sample, respectively.
- Eluted with 50μL Elution Buffer.
Spec’d on Roberts Lab NanoDrop1000 (ThermoFisher) and stored temporarily at 4C to avoid freeze-thawing before sending off for sequencing.
Yields are good (~6μg for the adductor muscle and ~10μg for the mantle).
The adductor muscle sample looks pretty good (perfect 260/280 ratio and solid 260/230 ratio), while the mantle sample looks OK (good 260/280 ratio, tolerable 260/230 ratio). Will run samples on gel to assess gDNA integrity.