Tag Archives: E.Z.N.A. Mollusc Kit

Genomic DNA Isolation – Olympia oyster adductor musle & mantle

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.

 

Results:

 

 

 

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.

 

Genomic DNA Isolation – Geoduck Adductor Muscle & Foot

Isolated gDNA from Panopea generosa (geoduck) adductor muscle & foot samples collected by Brent & Steven on 20150811 using the E.Z.N.A Mollusc DNA Kit (Omega Bio-Tek) according to the manufacturer’s protocol, with the following adjustments:

  • 41.8mg of adductor muscle
  • 30.0mg of foot used
  • 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 265μL of MBL Buffer
  • Added 514μL of 100% EtOH.
  • Eluted with 75μL Elution Buffer.

Spec’d on Roberts Lab NanoDrop1000 (ThermoFisher) and stored temporarily at 4C to avoid freeze-thawing before sending off for sequencing next week.

 

Results:

 

 

 

Yields are good (~7ug).

Quality (260/280 ratios) looks great for both samples.

260/230 ratio not very good for adductor muscle, but perfect for foot tissue.

Will run samples on gel to assess gDNA integrity.