2022-11-02 Barott and Brown Pdam RNA and DNA Extractions
RNA and DNA Extractions for Brown/Barott Pdam Project, November 2, 2022
Protocol Link
Extraction of 12 samples from Pocillopora damicornis (Pdam) from Heron Island in RNAlater March 2021 (Kristen Brown, Postdoc in Katie Barott lab)
Samples
Notes
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Continuting processing the Barott and Brown Pdam samples using the protocol detailed on 10/27/22.
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For my records: Took new Proteinase K, RNA/DNA Prep Buffer, and RNA/DNA Wash Buffer from the kit labelled 1/2 rec’d 10/4/22.
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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: 104.76 (S1) & 23344.85 (S2)
| Sample # | DNA_QBIT_1 | DNA_QBIT_2 | DNA_QBIT_AVG | Extraction.Notes |
|---|---|---|---|---|
| RS1B | 23.4 | 23 | 23.2 | Bead beat for 1 mins |
| RS2B | 7.74 | 7.6 | 7.67 | Bead beat for 2 mins |
| RS3B | 11.9 | 11.8 | 11.85 | Bead beat for 2 mins |
| RS8B | 22.6 | 22.6 | 22.6 | Bead beat for 1 mins |
| RS14C | 6.56 | 6.42 | 6.49 | Bead beat for 2 mins |
| RS15D | 9.62 | 9.5 | 9.56 | Bead beat for 2 mins |
| RF13B | 10.6 | 10.5 | 10.55 | Bead beat for 1 mins |
| RF14B | 11.1 | 11 | 11.05 | Bead beat for 1 mins |
| RF15D | 8.86 | 8.72 | 8.79 | Bead beat for 2 mins |
| RF19B | 23.8 | 23 | 23.4 | Bead beat for 1 mins |
| RF22B | 12.5 | 12.3 | 12.4 | Bead beat for 2.5 mins |
| RF24B | 24.4 | 24 | 24.2 | Bead beat for 1 mins |
RNA Standards: 417.37 (S1) & 9558.32 (S2)
| Sample # | RNA_QBIT_1 | RNA_QBIT_2 | RNA_QBIT_AVG | Extraction.Notes |
|---|---|---|---|---|
| RS1B | 22 | 22 | 22 | Bead beat for 1 mins |
| RS2B | 12 | 12.2 | 12.1 | Bead beat for 2 mins |
| RS3B | 12 | 12.2 | 12.1 | Bead beat for 2 mins |
| RS8B | 24.2 | 23.8 | 24 | Bead beat for 1 mins |
| RS14C | nd | nd | 0 | Bead beat for 2 mins |
| RS15D | nd | nd | 0 | Bead beat for 2 mins |
| RF13B | 12 | 12.4 | 12.2 | Bead beat for 1 mins |
| RF14B | nd | nd | 0 | Bead beat for 1 mins |
| RF15D | nd | nd | 0 | Bead beat for 2 mins |
| RF19B | 25.6 | 25.4 | 25.5 | Bead beat for 1 mins |
| RF22B | 13.6 | 14.2 | 13.9 | Bead beat for 2.5 mins |
| RF24B | 26 | 25.6 | 25.8 | Bead beat for 1 mins |
DNA and RNA Quality Check: Gel, using Kristina’s gel protocol at the bottom of this page.
Note: Gel was only run for 40 minutes due to time constraints so is a little smushed-looking but all lanes look great!
Next steps
Continuing on with the rest of this sample using this protocol!
Written on November 2, 2022