DNA Isolation and Quantification - C.gigas Diploid (Ronit) and Triploid (Nisbet)

Isolated some gDNA from the triploid Nisbet oysters we received on 20200218 and one of Ronit’s diploid ctenidia samples (Google Sheet) using the E.Z.N.A. Mollusc DNA Kit (Omega). See the “Results” section for sample info.

Samples were pulverized under liquid nitrogen (LN2). Samples were not weighed beforehand; simply “eye-balled” pieces to get close to ~30 - 50mg. Manufacturer’s protocol (PDF) was followed with the following notes/changes:

  • Proteinase K was not from kit (seem to be out). Instead was ThermoFisher brand (stored in -20C) with a concentration of ~20mg/mL and expiration date of September 2019.

  • eluted in 100uL of elution buffer

DNA was quantified using the Roberts Lab Qubit 3.0 using the dsDNA BR Assay (Invitrogen) and 1uL of each sample.

Resulting DNA was stored in Sam’s gDNA Box #3, Positions I4 - I9


Qubit results (Google Sheet):

Sample Original sample conc. (ng/uL) Elution volume (uL) Yield (ng)
Nisbet #1 adductor 50.4 100 5040
Nisbet #1 ctenidia 80.2 100 8020
Nisbet #1 gonad 14.8 100 1480
Nisbet #1 mantle 18.8 100 1880
Ronit D05 C2 89.4 100 8940
Ronit D04 C 97.0 100 9700