RNA and DNA Extractions for Brown/Barott Pdam Project, November 18, 2022

Protocol Link

Re-Extraction of 12 samples from Pocillopora damicornis (Pdam) from Heron Island in RNAlater March 2021 (Kristen Brown, Postdoc in Katie Barott lab)

Samples

Notes

• Continuting processing the Barott and Brown Pdam samples using the protocol detailed on 10/27/22.

• For these re-extractions, I had high-quality DNA and RNA for each sample but just not enough. I will proceed by adding 1-2 fragments depending on size to the original bead tube for each sample along with 400 uL of DNA/RNA shield that was previously removed. Then I will bead-beat as usual and continue as usual.

• See bead-beating notes below with Qubit values.

Qubit Results

• Used Broad range dsDNA and RNA Qubit Protocol
• All samples read twice, standard only read once

DNA Standards: 199.11 (S1) & 22469.88 (S2)

Sample #	DNA_QBIT_1	DNA_QBIT_2	DNA_QBIT_AVG
RS2B	37	36.2	36.6
RS2C	21	20.8	20.9
RS3B	40.3	40	40.15
RS6D	17.7	17.5	17.6
RS7B	47.2	46.6	46.9
RS14C	39.4	38.2	38.8
RS15D	28.4	28.4	28.4
RF13B	60.4	60.4	60.4
RF14B	75.6	75.4	75.5
RF15D	58.4	58.4	58.4
RF22B	32.4	32.8	32.6
RF25A	60.4	60.2	60.3

RNA Standards: 421.54 (S1) & 9880.18 (S2)

Sample #	RNA_QBIT_1	RNA_QBIT_2	RNA_QBIT_AVG
RS2B	30.2	29.8	30
RS2C	25	24.6	24.8
RS3B	38.4	38.4	38.4
RS6D	19	18.8	18.9
RS7B	37.8	37.8	37.8
RS14C	37.4	37.4	37.4
RS15D	23.4	23.8	23.6
RF13B	45.8	46.2	46
RF14B	65.2	65.4	65.3
RF15D	45.4	45.2	45.3
RF22B	35.6	35.8	35.7
RF25A	59.6	59	59.3

DNA and RNA Quality Check: Gel, using Kristina’s gel protocol at the bottom of this page.

Dilutung new ladder: 4 uL water, 1 uL loading dye, 1 uL ladder

Next steps

All done with these samples!