RNA and DNA Extractions for Brown/Barott Pdam Project, November 8, 2022

Protocol Link

Extraction of 18 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.

- Had much higher qubit values here because I started putting two of the tiny fragments into the bead beat tubes instead of one. Mostly this meant I only had to bead beat most samples for 1 minute instead of 2 to get a dark colored solution and resulted in great RNA! But I think the previous samples are great too, just slightly less in concentration.

• 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: 195.49 (S1) & 22843.75 (S2)

Sample #	DNA_QBIT_1	DNA_QBIT_2	DNA_QBIT_AVG
RS1C	9.58	9.36	9.47
RS2C	7.48	7.42	7.45
RS3D	12	11.8	11.9
RS7C	21.2	21.2	21.2
RS9C	27.2	26.8	27
RS11B	14.3	14	14.15
RS12C	18.9	19	18.95
RS13A	33.2	32.2	32.7
RS15B	15.8	15.5	15.65
RF13D	24.4	24.2	24.3
RD14C	11.8	11.7	11.75
RF16C	16.9	16.7	16.8
RF17D	24.4	24.4	24.4
RF18B	13.2	13.3	13.25
RF20B	15.2	15.3	15.25
RF22C	18.4	18.3	18.35
RF24D	14.8	14.7	14.75
RF25C	20.8	20.8	20.8

RNA Standards: 413.58 (S1) & 8989.92 (S2)

Sample #	RNA_QBIT_1	RNA_QBIT_2	RNA_QBIT_AVG	Bead Beat Time (mins)
RS1C	18.2	18.6	18.4	2
RS2C	11	11.4	11.2	2
RS3D	23.8	24.8	24.3	1
RS7C	20.4	20.6	20.5	2
RS9C	44.8	45.2	45	1
RS11B	23.3	22.8	23.05	1
RS12C	24.8	24.4	24.6	2
RS13A	42.2	41.8	42	1
RS15B	19.4	20	19.7	1
RF13D	33	32.8	32.9	1
RF14C	19	18.6	18.8	1
RF16C	26.4	26.6	26.5	2
RF17D	34.6	35.2	34.9	1
RF18B	27.2	27.4	27.3	1
RF20B	23.4	23.2	23.3	1
RF22C	29.2	29.2	29.2	1
RF24D	23.4	23.4	23.4	1
RF25C	28	28	28	1

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

Next steps

Continuing on with the rest of this sample using this protocol!