Samples from yesterday were centrifuged 30mins, 4000g, 4C (fixed angle rotor).

Supernatant was removed and pellets were dried (inverted tubes for 30mins).

Pellets were resuspended in 200uL H2O.

Data spreadsheet is here: 20131125 – Oyster Glycogen Assay

Finally located the remaining half of Emma’s samples. These had already been freeze dried AND pulverized! So, I just had to weigh out ~half of each sample for the glycogen assay.

Glycogen Assay:

Samples were placed in 3mL of 15% trichloracetic acid (TCA), vortexed and incubated for 1hr @ 4C.

Samples were spun 10mins, 3,000g, at 4C (in bucket rotor).

500uL of supernatant was transferred to 4mL of 100% EtOH, vortexed and incubated O/N @ 4C.

Will continue processing tomorrow.

Data spreadsheet is here: 20131125 – Oyster Glycogen Assay

Samples from yesterday were centrifuged 30mins, 4000g, 4C (fixed angle rotor).

Supernatant was removed and pellets were dried (inverted tubes for 30mins).

Pellets were resuspended in 200uL H2O and stored @ -20C.

Data spreadsheet is here: 20131125 – Oyster Glycogen Assay

Samples were previously freeze dried overnight and stored @ -20C. To maximize sample homogeneity and, thus, increase accuracy of both assays, all samples were mechanically pulverized in their existing tubes. Approximately half of each sample was weighed and used for the glycogen assay. The remainder of each sample was stored @ -20C.

Glycogen Assay:

Samples were placed in 3mL of 15% trichloracetic acid (TCA), vortexed and incubated for 1hr @ 4C.

Samples were spun 10mins, 3,000g, at 4C (in bucket rotor).

500uL of supernatant was transferred to 4mL of 100% EtOH, vortexed and incubated O/N @ 4C.

Will continue processing tomorrow.

Data spreadsheet is here: 20131125 – Oyster Glycogen Assay