2023-05-09 Project Summary, 2022-2023 RNA and DNA Extractions for ENCORE MDEC Project
Project Summary: 2022-2023 RNA and DNA Extractions for ENCORE MDEC Project
This is the project summary for RNA and DNA extractions as well as details for the preparation of two different RNA sequencing submissions: 1) One pooled library (from 4 samples) for RNAseq for the creation of the reference transcriptome; and 2) Submission of all MDEC RNA samples for TagSeq sequencing for gene expression. The samples were collected in August 2022 as part of the ENCORE project in the TPC-PI experiment. This post and project is focused only on Madracis decactis (MDEC). There are also samples of Diploria labyrinthiformis (DLAB), Montastraea cavernosa (MCAV), and Porites astreoides (PAST) shown in some of the extraction posts.
Outline of Content
- Project Summary: 2022-2023 RNA and DNA Extractions for ENCORE MDEC Project
- Outline of Content
- Extractions
- Extraction Protocol Link
- 2022-10-03: Extraction of 8 samples (2 MDEC)
- 2022-10-05: Extraction of 10 samples (2 MDEC)
- 2022-10-17: Extraction of 12 samples (3 MDEC)
- 2023-01-13: Extraction of 12 samples (3 MDEC)
- 2023-01-18: Extraction of 12 samples (4 MDEC)
- 2023-02-16: Extraction of 12 samples (3 MDEC)
- 2023-04-06: Extraction of 12 samples (3 MDEC)
- 2023-04-18: Extraction of 18 samples (4 MDEC)
- 2023-04-21: Extraction of 18 samples (4 MDEC)
- 2023-04-25: Extraction of 16 samples (9 MDEC)
- 2023-04-27: Extraction of 16 samples (1 MDEC)
- Reference Transcriptome: Pooling RNA Using Zymo Clean and Concentrate Kit (March 2, 2023)
- TagSeq: Concentrating Low-Concentration Samples to meet TagSeq Requeirements of 12 ng/uL
- TagSeq: Prep and submit RNA for ENCORE Project, May 8, 2023 (to be sent to UT Austin sometime this summer)
Extractions
Extraction Protocol Link
- Used Broad range dsDNA and RNA Qubit Protocol
- All samples read twice, standard only read once
- DNA and RNA Quality Check: Gel, using Kristina’s gel protocol at the bottom of this page.
2022-10-03: Extraction of 8 samples (2 MDEC)
Samples
Qubit Results
DNA Standards: 201.70 (S1) & 23899.02 (S2)
| colony_id | Species | Temp.Cat | DNA_QBIT_1 | DNA_QBIT_2 | DNA_QBIT_AVG |
|---|---|---|---|---|---|
| Mdec-A7 | Madracis decactis | 36 | 15 | 14.9 | 14.95 |
| Mdec-C6 | Madracis decactis | 26 | 6.24 | 6.08 | 6.16 |
RNA Standards: 414.70 (S1) & 9890.95 (S2)
| colony_id | Species | Temp.Cat | RNA_QBIT_1 | RNA_QBIT_2 | RNA_QBIT_AVG |
|---|---|---|---|---|---|
| Mdec-A7 | Madracis decactis | 36 | 18.4 | 18.2 | 18.3 |
| Mdec-C6 | Madracis decactis | 26 | 16.8 | 16.6 | 16.7 |
Gel
2022-10-05: Extraction of 10 samples (2 MDEC)
Samples
Qubit Results
DNA Standards: 198.66 (S1) & 24171.04 (S2)
| colony_id | Species | Temp.Cat | DNA_QBIT_1 | DNA_QBIT_2 | DNA_QBIT_AVG |
|---|---|---|---|---|---|
| Mdec-D4 | Madracis decactis | 16 | 5 | 4.78 | 4.89 |
| Mdec-E1 | Madracis decactis | 29 | 8.06 | 8.02 | 8.04 |
RNA Standards: 412.67 (S1) & 9635.27 (S2)
| colony_id | Species | Temp.Cat | RNA_QBIT_1 | RNA_QBIT_2 | RNA_QBIT_AVG |
|---|---|---|---|---|---|
| Mdec-D4 | Madracis decactis | 16 | 12.8 | 12.6 | 12.7 |
| Mdec-E1 | Madracis decactis | 29 | 22.2 | 21.8 | 22 |
Gel
2022-10-17: Extraction of 12 samples (3 MDEC)
Samples
Qubit Results
DNA Standards: 189.87 (S1) & 21953.74 (S2)
| colony_id | Species | Temp.Cat | DNA_QBIT_1 | DNA_QBIT_2 | DNA_QBIT_AVG |
|---|---|---|---|---|---|
| Mdec-A4 | Madracis decactis | 16 | 3.2 | 3.12 | 3.16 |
| Mdec-B5 | Madracis decactis | 22 | 7.18 | 7.2 | 7.19 |
| Mdec-F6 | Madracis decactis | 26 | 4.64 | 4.54 | 4.59 |
RNA Standards: 412.01 (S1) & 9743.89 (S2)
| colony_id | Species | Temp.Cat | RNA_QBIT_1 | RNA_QBIT_2 | RNA_QBIT_AVG |
|---|---|---|---|---|---|
| Mdec-A4 | Madracis decactis | 16 | nd | nd | 0 |
| Mdec-B5 | Madracis decactis | 22 | 14 | 14.2 | 14.1 |
| Mdec-F6 | Madracis decactis | 26 | 11.6 | 11.6 | 11.6 |
Gel
2023-01-13: Extraction of 12 samples (3 MDEC)
Samples
Qubit Results
DNA Standards: 189.48 (S1) & 20568.81 (S2)
| colony_id | Species | Temp.Cat | DNA_QBIT_1 | DNA_QBIT_2 | DNA_QBIT_AVG |
|---|---|---|---|---|---|
| Mdec-D6 | Madracis decactis | 26 | 2.68 | 2.68 | 2.68 |
| Mdec-E3 | Madracis decactis | 33 | 5 | 4.9 | 4.95 |
| Mdec-F1 | Madracis decactis | 29 | 13.9 | 14 | 13.95 |
RNA Standards: 409.92 (S1) & 9037.28 (S2)
| colony_id | Species | Temp.Cat | RNA_QBIT_1 | RNA_QBIT_2 | RNA_QBIT_AVG |
|---|---|---|---|---|---|
| Mdec-D6 | Madracis decactis | 26 | 11.6 | 11.4 | 11.5 |
| Mdec-E3 | Madracis decactis | 33 | 20.8 | 21.2 | 21 |
| Mdec-F1 | Madracis decactis | 29 | 53.8 | 53.6 | 53.7 |
Gel
2023-01-18: Extraction of 12 samples (4 MDEC)
Samples
Qubit Results
DNA Standards: 189.16 (S1) & 20760.08 (S2)
| colony_id | Species | Temp.Cat | DNA_QBIT_1 | DNA_QBIT_2 | DNA_QBIT_AVG |
|---|---|---|---|---|---|
| Mdec-B1 | Madracis decactis | 29 | 5.08 | 4.86 | 4.97 |
| Mdec-C3 | Madracis decactis | 33 | 4.34 | 4.34 | 4.34 |
| Mdec-E7 | Madracis decactis | 36 | 26.8 | 26.6 | 26.7 |
| Mdec-F4 | Madracis decactis | 16 | 7.4 | 7.32 | 7.36 |
RNA Standards: 414.76 (S1) & 9063.17 (S2)
| colony_id | Species | Temp.Cat | RNA_QBIT_1 | RNA_QBIT_2 | RNA_QBIT_AVG |
|---|---|---|---|---|---|
| Mdec-B1 | Madracis decactis | 29 | 13 | 13.6 | 13.3 |
| Mdec-C3 | Madracis decactis | 33 | 12.4 | 12.4 | 12.4 |
| Mdec-E7 | Madracis decactis | 36 | 26.2 | 26 | 26.1 |
| Mdec-F4 | Madracis decactis | 16 | 22.2 | 22 | 22.1 |
Gel
2023-02-16: Extraction of 12 samples (3 MDEC)
Samples
Qubit Results
DNA Standards: 167.18 (S1) & 16640.38 (S2)
| colony_id | Species | Temp.Cat | DNA_QBIT_1 | DNA_QBIT_2 | DNA_QBIT_AVG |
|---|---|---|---|---|---|
| Mdec-C2 | Madracis decactis | 40 | 18.5 | 18.5 | 18.5 |
| Mdec-D7 | Madracis decactis | 36 | 52 | 52 | 52 |
| Mdec-F7 | Madracis decactis | 36 | 23.2 | 23 | 23.1 |
RNA Standards: 385.96 (S1) & 8014.30 (S2)
| colony_id | Species | Temp.Cat | RNA_QBIT_1 | RNA_QBIT_2 | RNA_QBIT_AVG |
|---|---|---|---|---|---|
| Mdec-C2 | Madracis decactis | 40 | 33.2 | 32.8 | 33 |
| Mdec-D7 | Madracis decactis | 36 | 37.8 | 37.4 | 37.6 |
| Mdec-F7 | Madracis decactis | 36 | 10.6 | 0 | 5.3 |
Gel
2023-04-06: Extraction of 12 samples (3 MDEC)
Samples
Qubit Results
DNA Standards: 197.48 (S1) & 22870.14 (S2)
| colony_id | Species | Temp.Cat | DNA_QBIT_1 | DNA_QBIT_2 | DNA_QBIT_AVG |
|---|---|---|---|---|---|
| Mdec-A5 | Madracis decactis | 22 | 5.22 | 5.36 | 5.29 |
| Mdec-D3 | Madracis decactis | 33 | 6.72 | 6.76 | 6.74 |
| Mdec-E6 | Madracis decactis | 26 | 4.96 | 5 | 4.98 |
RNA Standards: 406.98 (S1) & 8665.72 (S2)
| colony_id | Species | Temp.Cat | RNA_QBIT_1 | RNA_QBIT_2 | RNA_QBIT_AVG |
|---|---|---|---|---|---|
| Mdec-A5 | Madracis decactis | 22 | 23.2 | 23.6 | 23.4 |
| Mdec-D3 | Madracis decactis | 33 | 31 | 31 | 31 |
| Mdec-E6 | Madracis decactis | 26 | 22.2 | 22 | 22.1 |
Mdec-D5 was also extracted this day, but was re-extracted on 2023-04-27 to improve yield. Only the re-extracted sample was sent for sequencing (TagSeq).
Gel
2023-04-18: Extraction of 18 samples (4 MDEC)
Samples
Qubit Results
DNA Standards: 191.24 (S1) & 21938.4 (S2)
| colony_id | Species | Temp.Cat | DNA_QBIT_1 | DNA_QBIT_2 | DNA_QBIT_AVG |
|---|---|---|---|---|---|
| Mdec-B6 | Madracis decactis | 26 | 6.22 | 6.36 | 6.29 |
| Mdec-B7 | Madracis decactis | 36 | 16.5 | 16.7 | 16.6 |
| Mdec-D1 | Madracis decactis | 29 | 8.4 | 8.38 | 8.39 |
| Mdec-F5 | Madracis decactis | 22 | 5.9 | 5.98 | 5.94 |
RNA Standards: 422.04 (S1) & 8600.79 (S2)
| colony_id | Species | Temp.Cat | RNA_QBIT_1 | RNA_QBIT_2 | RNA_QBIT_AVG |
|---|---|---|---|---|---|
| Mdec-B6 | Madracis decactis | 26 | 18.8 | 19 | 18.9 |
| Mdec-B7 | Madracis decactis | 36 | 14.8 | 15 | 14.9 |
| Mdec-D1 | Madracis decactis | 29 | 26.6 | 28 | 27.3 |
| Mdec-F5 | Madracis decactis | 22 | 15.2 | 15 | 15.1 |
Gel
2023-04-21: Extraction of 18 samples (4 MDEC)
Samples
The tube for MDEC F2 has the writing MDEC C2 on the side. However, there is also a complete MDEC C2 tube (labelled as such on side and cap) in the box and no other MDEC F2.
Continuing on with the rest of the ENCORE extractions.
Qubit Results
DNA Standards: 192.20 (S1) & 21884.44 (S2)
| colony_id | Species | Temp.Cat | DNA_QBIT_1 | DNA_QBIT_2 | DNA_QBIT_AVG |
|---|---|---|---|---|---|
| Mdec-B3 | Madracis decactis | 33 | 5.72 | 5.74 | 5.73 |
| Mdec-C4 | Madracis decactis | 16 | 8.4 | 8.38 | 8.39 |
| Mdec-E5 | Madracis decactis | 22 | 7.56 | 7.52 | 7.54 |
| Mdec-F2 | Madracis decactis | 40 | 15.5 | 15.8 | 15.65 |
RNA Standards: 417.49 (S1) & 8557.82 (S2)
| colony_id | Species | Temp.Cat | RNA_QBIT_1 | RNA_QBIT_2 | RNA_QBIT_AVG |
|---|---|---|---|---|---|
| Mdec-B3 | Madracis decactis | 33 | 14.4 | 14.4 | 14.4 |
| Mdec-C4 | Madracis decactis | 16 | 25.8 | 25.4 | 25.6 |
| Mdec-E5 | Madracis decactis | 22 | 28.4 | 28 | 28.2 |
| Mdec-F2 | Madracis decactis | 40 | 28.4 | 27.8 | 28.1 |
Gel
2023-04-25: Extraction of 16 samples (9 MDEC)
Samples
Qubit Results
DNA Standards: 198.48 (S1) & 22919.01 (S2)
| colony_id | Species | Temp.Cat | DNA_QBIT_1 | DNA_QBIT_2 | DNA_QBIT_AVG |
|---|---|---|---|---|---|
| Mdec-A1 | Madracis decactis | 29 | 2.88 | 2.98 | 2.93 |
| Mdec-A2 | Madracis decactis | 40 | 28.4 | 28.4 | 28.4 |
| Mdec-A3 | Madracis decactis | 33 | 6.26 | 6.38 | 6.32 |
| Mdec-B4 | Madracis decactis | 16 | 4.4 | 4.46 | 4.43 |
| Mdec-C1 | Madracis decactis | 29 | 8.52 | 8.6 | 8.56 |
| Mdec-C7 | Madracis decactis | 36 | 19.6 | 19.6 | 19.6 |
| Mdec-D2 | Madracis decactis | 40 | 22 | 22.4 | 22.2 |
| Mdec-E2 | Madracis decactis | 40 | 11.4 | 11.5 | 11.45 |
| Mdec-F3 | Madracis decactis | 33 | 4.72 | 4.8 | 4.76 |
RNA Standards: 423.98 (S1) & 8652.06 (S2)
| colony_id | Species | Temp.Cat | RNA_QBIT_1 | RNA_QBIT_2 | RNA_QBIT_AVG |
|---|---|---|---|---|---|
| Mdec-A1 | Madracis decactis | 29 | 6.9 | 6.6 | 6.75 |
| Mdec-A2 | Madracis decactis | 40 | 22.6 | 23 | 22.8 |
| Mdec-A3 | Madracis decactis | 33 | 15.8 | 15.6 | 15.7 |
| Mdec-B4 | Madracis decactis | 16 | 15 | 15.6 | 15.3 |
| Mdec-C1 | Madracis decactis | 29 | 26.2 | 26.4 | 26.3 |
| Mdec-C7 | Madracis decactis | 36 | 21.2 | 21 | 21.1 |
| Mdec-D2 | Madracis decactis | 40 | 21.6 | 21.8 | 21.7 |
| Mdec-E2 | Madracis decactis | 40 | 17.4 | 17.6 | 17.5 |
| Mdec-F3 | Madracis decactis | 33 | 15.4 | 16.2 | 15.8 |
Gel
2023-04-27: Extraction of 16 samples (1 MDEC)
Samples
Re-extractions:
Qubit Results
DNA Standards: 194.09 (S1) & 22253.03 (S2)
| colony_id | Species | Temp.Cat | DNA_QBIT_1 | DNA_QBIT_2 | DNA_QBIT_AVG |
|---|---|---|---|---|---|
| Mdec-D5 | Madracis decactis | 22 | 2.78 | 2.82 | 2.8 |
RNA Standards: 414.64 (S1) & 8566.59 (S2)
| colony_id | Species | Temp.Cat | RNA_QBIT_1 | RNA_QBIT_2 | RNA_QBIT_AVG |
|---|---|---|---|---|---|
| Mdec-D5 | Madracis decactis | 22 | 14.8 | 13.8 | 14.3 |
Gel
Reference Transcriptome: Pooling RNA Using Zymo Clean and Concentrate Kit (March 2, 2023)
For the purpose of creating a reference transcriptome, I made a pool of RNA from multiple samples to capture genetic diversity and treatment diversity in our reference transcriptome. The four samples are all from different genotypes and were all exposed to different temperatures (16, 22, 29, and 36 ºC). We will then send these three pools to Azenta for RNA sequencing. We used the Zymo RNA Clean and Concentrate Kit on the pools to concentrate the pooled RNA to the recommended concentration required by Azenta, >2 µg @ >50 ng/µL.
Samples
| colony_id | Temp.Cat | Extraction.Date | RNA_Qubit_AVG (ng/µL) |
|---|---|---|---|
| Mdec-F4 | 16 | 20230118 | 22.1 |
| Mdec-B5 | 22 | 20221017 | 14.1 |
| Mdec-E1 | 29 | 20221005 | 22 |
| Mdec-A7 | 36 | 20221003 | 18.3 |
Protocol
Take 40 µL per sample (4 samples total) and transfer into a new tube. Mix together, final volume in the tube will be 160 µL. This is pooling 4 tubes into 1, and this one tube will be all you work with for the clean and concentrate kit.
Then, follow the protocol for the Zymo Clean and Concentrate kit, eluting in 25 µL of DNAse/RNAse free water.
- Since each sample is 160 uL, add 320 uL of RNA binding buffer.
- Add an equal volume (480 uL) of 100% ethanol and mix.
- Transfer the sample to the Zymo-Spin™ IC Column in a Collection Tube and centrifuge. Discard the flow-through.
- Add 400 µl RNA Prep Buffer to the column and centrifuge. Discard the flow-through.
- Add 700 µl RNA Wash Buffer to the column and centrifuge. Discard the flow-through.
- Add 400 µl RNA Wash Buffer to the column and centrifuge for 1 minute ensure complete removal of the wash buffer. Carefully, transfer the column into a RNase-free tube (not provided).
- Add 25 µl DNase/RNase-Free Water directly to the column matrix and centrifuge.
Qubit Results
- Used Broad range dsDNA and RNA Qubit Protocol
- All samples read twice, standard only read once
RNA Standards: 416.13 (S1) & 9020.64 (S2)
| colony_id | Species | RNA_QBIT_1 | RNA_QBIT_2 | AVG (ng/uL) | Volume (uL), after Qubit | Final Amount RNA (ng) |
|---|---|---|---|---|---|---|
| Mdec Pool | Madracis decactis | 80.4 | 78.4 | 79.4 | 24 | 1905.6 |
Way over the minimum amount of 500 ng and well above the recommened concentration of 50 ng/uL. Ready for sequencing.
TagSeq: Concentrating Low-Concentration Samples to meet TagSeq Requeirements of 12 ng/uL
Follow the protocol for the Zymo Clean and Concentrate kit, eluting in 25 µL of DNAse/RNAse free water.
Base volume: 90 uL
- Since each sample is 90 uL, add 180 uL (2 x 90) of RNA binding buffer.
- Add an equal volume (270 uL (3 x 90)) of 100% ethanol and mix.
- Transfer the sample to the Zymo-Spin™ IC Column in a Collection Tube and centrifuge. Discard the flow-through.
- Add 400 µl RNA Prep Buffer to the column and centrifuge. Discard the flow-through.
- Add 700 µl RNA Wash Buffer to the column and centrifuge. Discard the flow-through.
- Add 400 µl RNA Wash Buffer to the column and centrifuge for 1 minute ensure complete removal of the wash buffer. Carefully, transfer the column into a RNase-free tube (not provided).
- Add 25 µl DNase/RNase-Free Water directly to the column matrix and centrifuge.
| Extraction.Date | Extraction.Notes | Volume in Tube currently | colony_id |
|---|---|---|---|
| 20230425 | 90 | Mdec-A1 | |
| 20221017 | 90 | Mdec-A4 | |
| 20230113 | 90 | Mdec-D6 |
Qubit Results
- Used High Sensitivity range dsDNA and RNA Qubit Protocol
- All samples read twice, standard only read once
RNA Standards: 33.21 (S1) & 438.03 (S2)
| colony_id | RNA_QBIT_AVG | Volume in Tube currently | Amount RNA in whole tube |
|---|---|---|---|
| Mdec-A1 | 13.3 | 24 | 319.2 |
| Mdec-A4 | 15.85 | 24 | 380.4 |
| Mdec-D6 | 15.8 | 24 | 379.2 |
All are now above 12 ng/uL and ready for plate prep!
TagSeq: Prep and submit RNA for ENCORE Project, May 8, 2023 (to be sent to UT Austin sometime this summer)
Dilute extracted RNA to 12 ug/uL in 25 uL and consolidate into one plate for TagSeq. This plate prep guide is based off of Dr. Kevin Wong’s protocol/example here and my previous post here.
Samples
All DLAB, MDEC, and MCAV – 119 samples.
| colony_id | RNA_QBIT_AVG | Amount to dilute, final concentration of 12 ng/uL in 25 uL | Amount of water | Volume in Tube currently | Amount left in tube after this | Plate Map Order | Plate Wells |
|---|---|---|---|---|---|---|---|
| Dlab-F1 | 15.9 | 19 | 6 | 40 | 21 | 1 | Plate1_A1 |
| Mdec-D4 | 12.7 | 24 | 1 | 90 | 66 | 2 | Plate1_B1 |
| Mcav-E1 | 27.3 | 11 | 14 | 50 | 39 | 3 | Plate1_C1 |
| Dlab-A2 | 17.4 | 17 | 8 | 35 | 18 | 4 | Plate1_D1 |
| Mdec-A5 | 23.4 | 13 | 12 | 90 | 77 | 5 | Plate1_E1 |
| Mcav-B5 | 18.1 | 17 | 8 | 24 | 7 | 6 | Plate1_F1 |
| Dlab-C5 | 13.4 | 22 | 3 | 40 | 18 | 7 | Plate1_G1 |
| Dlab-A1 | 12.6 | 24 | 1 | 90 | 66 | 8 | Plate1_H1 |
| Mdec-A7 | 18.3 | 16 | 9 | 50 | 34 | 9 | Plate1_A2 |
| Dlab-F2 | 18 | 17 | 8 | 35 | 18 | 10 | Plate1_B2 |
| Dlab-C7 | 17.15 | 17 | 8 | 24 | 7 | 11 | Plate1_C2 |
| Mcav-E2 | 27.4 | 11 | 14 | 90 | 79 | 12 | Plate1_D2 |
| Mdec-D5 | 14.3 | 21 | 4 | 40 | 19 | 13 | Plate1_E2 |
| Dlab-C6 | 13.5 | 22 | 3 | 40 | 18 | 14 | Plate1_F2 |
| Mcav-B6 | 17 | 18 | 7 | 24 | 6 | 15 | Plate1_G2 |
| Dlab-A3 | 13.55 | 22 | 3 | 24 | 2 | 16 | Plate1_H2 |
| Mcav-E3 | 19.9 | 15 | 10 | 90 | 75 | 17 | Plate1_A3 |
| Mcav-B7 | 23.3 | 13 | 12 | 90 | 77 | 18 | Plate1_B3 |
| Mdec-B1 | 13.3 | 23 | 2 | 90 | 67 | 19 | Plate1_C3 |
| Dlab-F3 | 19.1 | 16 | 9 | 24 | 8 | 20 | Plate1_D3 |
| Mdec-D6 | 15.8 | 19 | 6 | 24 | 5 | 21 | Plate1_E3 |
| Dlab-F4 | 18.9 | 16 | 9 | 24 | 8 | 22 | Plate1_F3 |
| Dlab-D1 | 12.3 | 24 | 1 | 50 | 26 | 23 | Plate1_G3 |
| Dlab-A4 | 13.5 | 22 | 3 | 50 | 28 | 24 | Plate1_H3 |
| Mdec-B3 | 14.4 | 21 | 4 | 90 | 69 | 25 | Plate1_A4 |
| Dlab-D2 | 21.2 | 14 | 11 | 40 | 26 | 26 | Plate1_B4 |
| Dlab-F5 | 14 | 21 | 4 | 24 | 3 | 27 | Plate1_C4 |
| Dlab-A5 | 15.65 | 19 | 6 | 24 | 5 | 28 | Plate1_D4 |
| Mcav-C1 | 18.5 | 16 | 9 | 90 | 74 | 29 | Plate1_E4 |
| Mdec-D7 | 37.6 | 8 | 17 | 90 | 82 | 30 | Plate1_F4 |
| Mcav-C2 | 14.9 | 20 | 5 | 90 | 70 | 31 | Plate1_G4 |
| Mcav-E4 | 12 | 25 | 0 | 90 | 65 | 32 | Plate1_H4 |
| Dlab-F6 | 25.5 | 12 | 13 | 24 | 12 | 33 | Plate1_A5 |
| Dlab-A6 | 12 | 25 | 0 | 40 | 15 | 34 | Plate1_B5 |
| Dlab-D3 | 12.2 | 25 | 0 | 90 | 65 | 35 | Plate1_C5 |
| Mcav-E5 | 17.45 | 17 | 8 | 24 | 7 | 36 | Plate1_D5 |
| Mdec-B4 | 15.3 | 20 | 5 | 90 | 70 | 37 | Plate1_E5 |
| Mdec-E1 | 22 | 14 | 11 | 50 | 36 | 38 | Plate1_F5 |
| Mcav-E6 | 17.25 | 17 | 8 | 24 | 7 | 39 | Plate1_G5 |
| Mcav-C3 | 21.6 | 14 | 11 | 90 | 76 | 40 | Plate1_H5 |
| Mcav-C4 | 12.2 | 25 | 0 | 90 | 65 | 41 | Plate1_A6 |
| Dlab-A7 | 17.55 | 17 | 8 | 24 | 7 | 42 | Plate1_B6 |
| Mdec-B5 | 16.9 | 18 | 7 | 50 | 32 | 43 | Plate1_C6 |
| Dlab-D4 | 12 | 25 | 0 | 40 | 15 | 44 | Plate1_D6 |
| Mcav-F1 | 34.4 | 9 | 16 | 90 | 81 | 45 | Plate1_E6 |
| Dlab-F7 | 18 | 17 | 8 | 40 | 23 | 46 | Plate1_F6 |
| Mdec-E2 | 17.5 | 17 | 8 | 90 | 73 | 47 | Plate1_G6 |
| Mdec-B6 | 18.9 | 16 | 9 | 90 | 74 | 48 | Plate1_H6 |
| Mdec-E3 | 21 | 14 | 11 | 90 | 76 | 49 | Plate1_A7 |
| Mcav-C5 | 14.8 | 20 | 5 | 23 | 3 | 50 | Plate1_B7 |
| Mcav-F2 | 24.1 | 12 | 13 | 90 | 78 | 51 | Plate1_C7 |
| Mcav-A1 | 13.3 | 23 | 2 | 90 | 67 | 52 | Plate1_D7 |
| Mdec-B7 | 14.9 | 20 | 5 | 90 | 70 | 53 | Plate1_E7 |
| Dlab-D5 | 14 | 21 | 4 | 40 | 19 | 54 | Plate1_F7 |
| Mdec-E5 | 28.2 | 11 | 14 | 90 | 79 | 55 | Plate1_G7 |
| Dlab-B1 | 12.35 | 24 | 1 | 24 | 0 | 56 | Plate1_H7 |
| Dlab-B2 | 19.4 | 15 | 10 | 40 | 25 | 57 | Plate1_A8 |
| Dlab-D6 | 13.4 | 22 | 3 | 24 | 2 | 58 | Plate1_B8 |
| Mdec-C1 | 26.3 | 11 | 14 | 90 | 79 | 59 | Plate1_C8 |
| Mcav-F3 | 12.1 | 25 | 0 | 90 | 65 | 60 | Plate1_D8 |
| Mdec-E6 | 22.1 | 14 | 11 | 90 | 76 | 61 | Plate1_E8 |
| Mcav-C6 | 14 | 21 | 4 | 40 | 19 | 62 | Plate1_F8 |
| Mcav-A2 | 17.2 | 17 | 8 | 90 | 73 | 63 | Plate1_G8 |
| Dlab-B3 | 18.5 | 16 | 9 | 35 | 19 | 64 | Plate1_H8 |
| Dlab-D7 | 19.6 | 15 | 10 | 40 | 25 | 65 | Plate1_A9 |
| Mdec-E7 | 26.1 | 11 | 14 | 90 | 79 | 66 | Plate1_B9 |
| Mdec-C2 | 33 | 9 | 16 | 90 | 81 | 67 | Plate1_C9 |
| Mcav-F4 | 18.2 | 16 | 9 | 40 | 24 | 68 | Plate1_D9 |
| Dlab-E1 | 16.2 | 19 | 6 | 40 | 21 | 69 | Plate1_E9 |
| Mcav-C7 | 17.1 | 18 | 7 | 90 | 72 | 70 | Plate1_F9 |
| Dlab-B4 | 12.5 | 24 | 1 | 40 | 16 | 71 | Plate1_G9 |
| Mcav-A3 | 17.2 | 17 | 8 | 90 | 73 | 72 | Plate1_H9 |
| Mcav-A4 | 24.1 | 12 | 13 | 90 | 78 | 73 | Plate1_A10 |
| Mdec-C3 | 12.4 | 24 | 1 | 90 | 66 | 74 | Plate1_B10 |
| Mcav-D1 | 14.5 | 21 | 4 | 90 | 69 | 75 | Plate1_C10 |
| Dlab-B5 | 16.05 | 19 | 6 | 24 | 5 | 76 | Plate1_D10 |
| Mcav-F5 | 18.55 | 16 | 9 | 24 | 8 | 77 | Plate1_E10 |
| Mdec-F1 | 53.7 | 6 | 19 | 90 | 84 | 78 | Plate1_F10 |
| Mcav-A5 | 17.1 | 18 | 7 | 24 | 6 | 79 | Plate1_G10 |
| Dlab-E2 | 25.1 | 12 | 13 | 40 | 28 | 80 | Plate1_H10 |
| Mcav-F6 | 15.3 | 20 | 5 | 40 | 20 | 81 | Plate1_A11 |
| Mcav-D2 | 15.9 | 19 | 6 | 90 | 71 | 82 | Plate1_B11 |
| Mdec-C4 | 25.6 | 12 | 13 | 90 | 78 | 83 | Plate1_C11 |
| Mcav-D3 | 19.4 | 15 | 10 | 90 | 75 | 84 | Plate1_D11 |
| Dlab-B6 | 15.4 | 19 | 6 | 40 | 21 | 85 | Plate1_E11 |
| Mdec-F2 | 28.1 | 11 | 14 | 90 | 79 | 86 | Plate1_F11 |
| Dlab-E3 | 15 | 20 | 5 | 40 | 20 | 87 | Plate1_G11 |
| Mcav-A6 | 15.1 | 20 | 5 | 90 | 70 | 88 | Plate1_H11 |
| Mcav-A7 | 23.5 | 13 | 12 | 50 | 37 | 89 | Plate1_A12 |
| Dlab-E4 | 14.1 | 21 | 4 | 24 | 3 | 90 | Plate1_B12 |
| Mcav-D4 | 16.9 | 18 | 7 | 90 | 72 | 91 | Plate1_C12 |
| Mdec-F3 | 15.8 | 19 | 6 | 90 | 71 | 92 | Plate1_D12 |
| Mdec-A1 | 13.3 | 23 | 2 | 24 | 1 | 93 | Plate1_E12 |
| Mcav-F7 | 15.3 | 20 | 5 | 24 | 4 | 94 | Plate1_F12 |
| Dlab-B7 | 13.7 | 22 | 3 | 40 | 18 | 95 | Plate1_G12 |
| Mdec-C6 | 16.7 | 18 | 7 | 90 | 72 | 96 | Plate1_H12 |
| Mcav-B1 | 14.8 | 20 | 5 | 90 | 70 | 97 | Plate2_A1 |
| Mdec-F4 | 22.1 | 14 | 11 | 50 | 36 | 98 | Plate2_B1 |
| Mdec-A2 | 22.8 | 13 | 12 | 90 | 77 | 99 | Plate2_C1 |
| Mcav-D5 | 16.4 | 18 | 7 | 50 | 32 | 100 | Plate2_D1 |
| Dlab-C2 | 17.3 | 17 | 8 | 90 | 73 | 101 | Plate2_E1 |
| Mdec-D1 | 27.3 | 11 | 14 | 90 | 79 | 102 | Plate2_F1 |
| Dlab-E5 | 14.4 | 21 | 4 | 40 | 19 | 103 | Plate2_G1 |
| Mdec-C7 | 21.1 | 14 | 11 | 90 | 76 | 104 | Plate2_H1 |
| Dlab-E6 | 12.25 | 23 | 2 | 23 | 0 | 105 | Plate2_A2 |
| Dlab-C3 | 14 | 21 | 4 | 40 | 19 | 106 | Plate2_B2 |
| Mcav-B2 | 27.1 | 11 | 14 | 90 | 79 | 107 | Plate2_C2 |
| Mdec-F5 | 15.1 | 20 | 5 | 90 | 70 | 108 | Plate2_D2 |
| Mdec-D2 | 21.7 | 14 | 11 | 90 | 76 | 109 | Plate2_E2 |
| Mcav-D6 | 19 | 16 | 9 | 24 | 8 | 110 | Plate2_F2 |
| Mdec-A3 | 15.7 | 19 | 6 | 90 | 71 | 111 | Plate2_G2 |
| Mdec-F6 | 14.5 | 21 | 4 | 90 | 69 | 112 | Plate2_H2 |
| Mdec-A4 | 15.85 | 19 | 6 | 24 | 5 | 113 | Plate2_A3 |
| Dlab-E7 | 17.8 | 17 | 8 | 90 | 73 | 114 | Plate2_B3 |
| Mdec-D3 | 31 | 10 | 15 | 90 | 80 | 115 | Plate2_C3 |
| Mcav-B4 | 17.1 | 18 | 7 | 50 | 32 | 116 | Plate2_D3 |
| Mcav-D7 | 16 | 19 | 6 | 90 | 71 | 117 | Plate2_E3 |
| Dlab-C4 | 13 | 23 | 2 | 35 | 12 | 118 | Plate2_F3 |
| Mdec-F7 | 21.5 | 14 | 11 | 90 | 76 | 119 | Plate2_G3 |
| Plate 1 | |||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Dlab-F1 | Mdec-A7 | Mcav-E3 | Mdec-B3 | Dlab-F6 | Mcav-C4 | Mdec-E3 | Dlab-B2 | Dlab-D7 | Mcav-A4 | Mcav-F6 | Mcav-A7 |
| Mdec-D4 | Dlab-F2 | Mcav-B7 | Dlab-D2 | Dlab-A6 | Dlab-A7 | Mcav-C5 | Dlab-D6 | Mdec-E7 | Mdec-C3 | Mcav-D2 | Dlab-E4 |
| Mcav-E1 | Dlab-C7 | Mdec-B1 | Dlab-F5 | Dlab-D3 | Mdec-B5 | Mcav-F2 | Mdec-C1 | Mdec-C2 | Mcav-D1 | Mdec-C4 | Mcav-D4 |
| Dlab-A2 | Mcav-E2 | Dlab-F3 | Dlab-A5 | Mcav-E5 | Dlab-D4 | Mcav-A1 | Mcav-F3 | Mcav-F4 | Dlab-B5 | Mcav-D3 | Mdec-F3 |
| Mdec-A5 | Mdec-D5 | Mdec-D6 | Mcav-C1 | Mdec-B4 | Mcav-F1 | Mdec-B7 | Mdec-E6 | Dlab-E1 | Mcav-F5 | Dlab-B6 | Mdec-A1 |
| Mcav-B5 | Dlab-C6 | Dlab-F4 | Mdec-D7 | Mdec-E1 | Dlab-F7 | Dlab-D5 | Mcav-C6 | Mcav-C7 | Mdec-F1 | Mdec-F2 | Mcav-F7 |
| Dlab-C5 | Mcav-B6 | Dlab-D1 | Mcav-C2 | Mcav-E6 | Mdec-E2 | Mdec-E5 | Mcav-A2 | Dlab-B4 | Mcav-A5 | Dlab-E3 | Dlab-B7 |
| Dlab-A1 | Dlab-A3 | Dlab-A4 | Mcav-E4 | Mcav-C3 | Mdec-B6 | Dlab-B1 | Dlab-B3 | Mcav-A3 | Dlab-E2 | Mcav-A6 | Mdec-C6 |
| Plate 2 | ||
|---|---|---|
| Mcav-B1 | Dlab-E6 | Mdec-A4 |
| Mdec-F4 | Dlab-C3 | Dlab-E7 |
| Mdec-A2 | Mcav-B2 | Mdec-D3 |
| Mcav-D5 | Mdec-F5 | Mcav-B4 |
| Dlab-C2 | Mdec-D2 | Mcav-D7 |
| Mdec-D1 | Mcav-D6 | Dlab-C4 |
| Dlab-E5 | Mdec-A3 | Mdec-F7 |
| Mdec-C7 | Mdec-F6 |
Prepped plates were covered with foil and placed in -80 until shipping:
