Kaitlyn’s notebook: Geoduck RNA extraction

RNA was isolated with a Quick-DNA/RNA Microprep Plus Kit by ZymoResearch according to the manufacturer’s protocol from geoduck samples .

For hemolymph, 150ul of sample was taken and 600ul of lysis buffer was added for prep, except for sample 27 which contained 120ul of sample and had 48-ul of lysis buffer added. All centrifuge steps were done at 16,000 rpm.

The on-column DNase step was done, and the elution volume is 15ul.

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.

The samples are stored in a box in the -80C freezer in 3, 3, 2, labelled “RNA isolations; geoduck 12/17”.

Sample RNA (ng/ul)
Star 11/15 47.2
Chewy 11/15 92.6
Star 11/21 170
Chewy 11/21 180
1 12.5
2 33.8
40 low
62 4.2
58 low
47 low
53 low
50 low
19 4.6
66 4.2
22 low
29 low
32 low
59 100
63 low
27 134
64 low
Standard 1 38.07
Standard 2 395.24

Samples are located in:

  • Box 8,1,1 (2014): Geo- 27, 31, 29
  • Box 8,1,3 (2015): Geo-  50, 58, 62, 19, 64, 40, 59, 47, 53, 63, 22, 66.
20191217_082151

Box 3,1,1 on left and Box 8,1,3 on right separated by Star and Chewy samples. Samples 1 and 2 and Star and Chewy samples are in box 5,3,1.

Sample Geo-30 H did not exist, although a Geo-30 did but it looked like gonad so I did not extract from it. 20191217_082036.jpg

Notes:

Sample 53 was dark green.