Kaitlyn’s notebook: Hemolymph RNA extractions 2

RNA was isolated with a Quick-DNA/RNA Microprep Plus Kit by ZymoResearch. The samples are stored in a box in the -80C freezer in 3, 3, 2, labelled “RNA isolations; geoduck 12/17”. Isolations were done according according to the manufacturer’s protocol on the following samples:

G-32 H
G-47 H
G-53 H
G-59 H
G-62 H
G-63 H
G-29 H
G-19 H
G-22 H
G-64 H
G-66 H
G-40 H
G-58 H
G-50 H

For hemolymph, 600ul of sample was taken and 2400ul of lysis buffer was added for prep. If 600ul of sample was not available, all of the sample was taken. The on-column DNase step was done, and the elution volume was 15ul.

Several columns were getting clogged from tissue in the sample.

Samples were quantified with the hsRNA Assay for Qubit according to manufacturer’s protocol. 1ul of sample was used and 199ul of working solution was used in each assay tube. All values were too low for quantification which was lickely caused by overloading the column, column clogging, and/or unknown technical error.