Appendix 2.
List of taxa from which RNA was extracted, the method used, and the Agilent 2100 Bioanalyzer quality indicators of the final extraction that was sequenced.a
| Option 1 | Option 2 | Option 3 | ||||||||||
| Family | Taxon | Material isolated (μg) | RINb | rRNA Ratio 26S/18S | Material isolated (μg) | RINb | rRNA Ratio 26S/18S | Material isolated (μg) | RINb | rRNA Ratio 26S/18S | Sequenced? | Voucher ID |
| Acanthaceae | Anisacanthus quadrifidus (Vahl) Nees | 40 | 6.4 | Missing | — | — | — | 46.1 | 3.9 | 1.2 | Option 1 | Soltis and Miles 2970 |
| Acanthaceae | Ruellia brittoniana Leonard | 43 | 7.6 | Missing | — | — | — | 29.2 | 2.4 | 0 | Option 1 | Soltis and Miles 2962 |
| Acanthaceae | Strobilanthes dyerianus | — | — | — | — | — | — | 25* | 4.9 | 2.7 | Option 3 | Soltis and Miles 2958 |
| Adoxaceae | Sambucus canadensis L. | 45.5 | 4.2 | 1.4 | — | — | — | — | — | — | Option 1 | Soltis and Miles 2955 |
| Amaryllidaceae | Sprekelia formosissima (L.) Herb. | — | — | — | 22.1 | 3.2 | 0.2 | — | — | — | Library construction failed | Garcia 4381 |
| Amaryllidaceae | Traubia modesta Ravenna | — | — | — | 40 | 6 | 1.6 | — | — | — | Option 2 | Garcia 3014 |
| Amaryllidaceae | Zephyranthes citrina Baker | — | — | — | 22 | 4.5 | 0.3 | — | — | — | Library construction failed | Garcia 4376 |
| Araliaceae | Hydrocotyle umbellata L. | — | — | — | — | — | — | 30* | 5.9 | 1.1 | Option 3 | Soltis and Miles 2959 |
| Asparagaceae | Disporopsis pernyi (Hua) Diels | 33.7 | 7.2 | 1.4 | — | — | — | — | — | — | Option 1 | Mike Heaney JMH2546 |
| Asparagaceae | Nolina bigelovii (Torr.) S. Watson | 33.45 | 7.9 | 1.5 | — | — | — | — | — | — | Option 1 | Mike Heaney JMH2984 |
| Asparagaceae | Peliosanthes minor Yamam. | 21.1 | 6.9 | 1.2 | — | — | — | — | — | — | Option 1 | Mike Heaney JMH2549 |
| Asteraceae | Cicerbita plumieri Kirschl. | 21 | 7.6 | 2.5 | — | — | — | — | — | — | Option 1 | Edward Schilling 11-48 |
| Asteraceae | Lactuca graminifolia Michx. | 54.85 | 7.6 | 2.4 | — | — | — | — | — | — | Option 1 | Edward Schilling LG-1 |
| Asteraceae | Tragopogon castellanus Levier | 19.3 | 6.3 | 1 | — | — | — | 36.7 | 2.1 | 0 | Option 1 | Soltis and Miles 2993 |
| Berberidaceae | Nandina domestica Thunb. | — | — | — | — | — | — | 30* | 5.7 | 1 | Option 3 | Soltis and Miles 2972 |
| Betulaceae | Alnus serrulata (Aiton) Willd. | — | — | — | — | — | — | 20* | 8.2 | 2.2 | Option 3 | Soltis and Miles 2964 |
| Bignoniaceae | Kigelia africana (Lam.) Benth. | 250.9 | 5.1 | 2.8 | — | — | — | — | — | — | Option 1 | Soltis and Miles 2992 |
| Bixaceae | Bixa orellana L. | — | — | — | — | — | — | 30* | 4.8 | 1.6 | Option 3 | Soltis and Miles 2985 |
| Cactaceae | Opuntia austrina Small | — | — | — | 7.7 | 8.2 | 1.4 | — | — | — | Low conc.; library not tried | Lucas Majure LCM_3450 |
| Cactaceae | Opuntia pusilla (Haw.) Haw. | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | All extractions failed | Lucas Majure LCM_753 |
| Calophyllaceae | Mammea americana L. | — | — | — | 50 | 6 | 1.6 | — | — | — | Option 2 | Soltis and Miles 3003 |
| Canellaceae | Canella winterana (L.) Gaertn. | 42.0 | 6.3 | 1.1 | — | — | — | 28.5 | 2.6 | 0.5 | Option 1 | Soltis and Miles 2995 |
| Chrysobalanaceae | Licania michauxii Prance | — | — | — | 22 | 7 | 2 | — | — | — | Option 2 | Soltis and Miles 2990 |
| Corynocarpaceae | Corynocarpus laevigatus J. R. Forst. & G. Forst. | 126.8 | 7.3 | Missing | — | — | — | 42.2 | 3.4 | 0.8 | Option 1 | Soltis and Miles 2986 |
| Didiereaceae | Alluaudiopsis marnieriana Rauh | 15.9 | 5.9 | Missing | — | — | — | 27.7 | 3.3 | 0.8 | Option 1 | Soltis and Miles 2981 |
| Fagaceae | Castanea pumila (L.) Mill. | — | — | — | — | — | — | 25* | 6.5 | 1.5 | Option 3 | Soltis and Miles 2977 |
| Gunneraceae | Gunnera chilensis Lam. | — | — | — | — | — | — | 25* | 6.7 | 1.6 | Option 3 | Soltis and Miles 2966 |
| Gunneraceae | Gunnera manicata Linden ex André | — | — | — | — | — | — | 81 | 5.6 | 1 | Option 3 | Soltis and Miles 2938 |
| Haemodoraceae | Lachnanthes caroliniana (Lam.) Dandy | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | All extractions failed | Soltis and Miles 2988 |
| Hydrangeaceae | Deutzia scabra Thunb. | — | — | — | 1.75 | 2.6 | 0 | 82.3 | 2.9 | 0.5 | Option 3 | Soltis and Miles 2965 |
| Hydrangeaceae | Deutzia scabra Thunb. | Low conc.; library not tried | Soltis and Miles 2965 | |||||||||
| Hydrangeaceae | Philadelphus inodorus L. | — | — | — | — | — | — | 35* | 4.9 | 1.1 | Option 3 | Soltis and Miles 2953 |
| Jubulaceae | Frullania sp. Raddi | — | — | — | 40 | 6.1 | 1.6 | — | — | — | Option 2 | Von Konrat 10021 |
| Kirkiaceae | Kirkia wilmsii Engl. | — | — | — | — | — | — | 13.8* | 6.9 | 0.7 | Option 3 | Soltis and Miles 2982 |
| Krameriaceae | Krameria lanceolata Torr. | — | — | — | — | — | — | 23.8 | 4.5 | 1.1 | Option 3 | Soltis and Miles 2991 |
| Lamiaceae | Pogostemon sp. Desf. | 39.35 | 7.5 | 1.6 | — | — | — | — | — | — | Option 1 | Grant Godden GGT4 |
| Lamiaceae | Poliomintha bustamanta B. L. Turner | — | — | — | 46 | 6 | 1.6 | — | — | — | Option 2 | Grant Godden GGT1 |
| Lamiaceae | Teucrium chamaedrys L. | 72.85 | 7.4 | 1.7 | — | — | — | — | — | — | Option 1 | Grant Godden GGT3 |
| Lamiaceae | Thymus vulgaris L. | 71.1 | 7.5 | 1.4 | — | — | — | — | — | — | Option 1 | Grant Godden GGT2 |
| Lauraceae | Cryptocarya alba (Molina) Looser | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | All extractions failed | Soltis and Miles 2998 |
| Lauraceae | Lindera benzoin (L.) Blume | 2.7 | 2.6 | 0 | 48.5 | 6 | 1.6 | 27.8 | 2.7 | 0.2 | Option 2 | Soltis and Miles 2968 |
| Lauraceae | Persea borbonia (L.) Spreng. | — | — | — | — | — | — | 18* | 5.3 | 1.2 | Option 3 | Soltis and Miles 2980 |
| Lentibulariaceae | Utricularia sp. L. | — | — | — | — | — | — | 30.9 | 4.3 | 1.2 | Option 3 | Soltis and Miles 2987 |
| Lythraceae | Lagerstroemia indica L. | — | — | — | — | — | — | 7 | 8 | 1.8 | Option 3 | Soltis and Miles 2971 |
| Lythraceae | Punica granatum L. | — | — | — | — | — | — | 20* | 7.5 | 1.9 | Option 3 | Soltis and Miles 2973 |
| Magnoliaceae | Michelia maudiae Dunn | — | — | — | — | — | — | 28.5* | 4.1 | 0.6 | Option 3 | Soltis and Miles 2954 |
| Malvaceae | Grewia occidentalis | 14.04 | 3.2 | 1 | Degraded | Missing | ||||||
| Meliaceae | Melia azedarach L. | 202 | 6.5 | 1.5 | — | — | — | — | — | — | Option 1 | Soltis and Miles 2961 |
| Muntingiaceae | Muntingia calabura L. | — | — | — | 21 | 8.1 | 1.7 | 122.8 | 2.9 | 0.3 | Option 2 | Soltis and Miles 2984 |
| Myricaceae | Myrica pumila (Michx.) Small | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | 14.5 | 3.1 | 1.3 | All extractions failed | Jackie Rice JDR9 |
| Oleaceae | Forestiera segregata (Jacq.) Krug & Urb. | 27.0 | 6.5 | Missing | — | — | — | 24.5 | 3.0 | 0.5 | Option 1 | Soltis and Miles 2969 |
| Orobanchaceae | Epifagus virginiana (L.) W. P. C. Barton | 10.9 | 3.1 | 0.6 | 38.3 | 7.2 | 1.6 | 46.5 | 2.1 | 0 | Option 2 | Soltis and Miles 2996 (Paul Manos 1800) |
| Phyllanthaceae | Bischofia javanica Blume | — | — | — | — | — | — | 10* | 7.8 | 3 | Option 3 | Soltis and Miles 2978 |
| Phytolaccaceae | Hilleria latifolia (Lam.) H. Walter | 24.1 | 5.8 | 0.5 | — | — | — | 35.6 | 2.7 | 0.1 | Option 1 | Soltis and Miles 2976 |
| Phytolaccaceae | Microtea debilis Sw. | 43.9 | 6.3 | 1 | — | — | — | 29.4 | 3.0 | 0.4 | Option 1 | Soltis and Miles 2997 |
| Phytolaccaceae | Rivina humilis L. | 19.45 | 6.6 | 1 | — | — | — | 28.42 | 2.8 | 0.4 | Option 1 | Soltis and Miles 2975 |
| Plantaginaceae | Antirrhinum majus L. | 64.0 | 5.4 | 0.5 | — | — | — | 50.9 | 4.1 | 0.8 | Option 1 | Soltis and Miles 2963 |
| Plantaginaceae | Bacopa caroliniana (Walter) B. L. Rob. | 26.0 | 7.8 | 1.4 | — | — | — | 28.4 | 3.5 | 1.5 | Option 1 | Soltis and Miles 2974 |
| Poaceae | Aristida stricta Michx. | 54.5 | 5.6 | Missing | — | — | — | — | — | — | Option 1 | Jackie Rice JDR10 |
| Poaceae | Uniola paniculata L. | 35.85 | 7.6 | 2.5 | — | — | — | — | — | — | Option 1 | Richard Hodel RGJH1001 |
| Podostemaceae | Podostemum sp. Michx. | — | — | — | 23.9 | 4.7 | 0.8 | 44.7 | 2.1 | 0 | Library construction failed | Soltis and Miles 2994 |
| Pteridaceae | Ceratopteris sp. Brongn. | — | — | — | Missing | Missing | Missing | — | — | — | Option 2 | Soltis and Miles 3005 |
| Pteridaceae | Pteris ensiformis Burm. f. | 28 | 8.4 | 1.6 | — | — | — | — | — | — | Option 1 | Soltis and Miles 3001 |
| Rhamnaceae | Rhamnus caroliniana Walter | 162 | 5.8 | 1.1 | — | — | — | 80 | 3.1 | 0.5 | Option 1 | Soltis and Miles 2952 |
| Rhamnaceae | Ziziphus jujuba Mill. | 0.15 | 2.7 | 0 | 45 | 6.5 | 1.6 | 25* | 3.6 | 0.6 | Option 2 | Soltis and Miles 2956 |
| Rosaceae | Agrimonia eupatoria L. | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | All extractions failed | Chase 38775 |
| Rosaceae | Amelanchier canadensis (L.) Medik. | — | — | — | 50 | 7 | 1.7 | — | — | — | Option 2 | Chase 38778 |
| Rosaceae | Aruncus dioicus (Walter) Fernald | — | — | — | 122 | 6.4 | 1.6 | — | — | — | Option 2 | Chase 38772 |
| Rosaceae | Cercocarpus ledifolius Nutt. | — | — | — | 40 | 7.1 | 2.5 | — | — | — | Option 2 | Chase 38780 |
| Rosaceae | Cotoneaster transcaucasicus Pojark. | — | — | — | 154 | 8.2 | 1.5 | — | — | — | Option 2 | Chase 38779 |
| Rosaceae | Dryas octopetala L. | — | — | — | 67 | 7.3 | 1.6 | — | — | — | Option 2 | Chase 38781 |
| Rosaceae | Geum quellyon Sweet | — | — | — | 78.2 | 7.7 | 1.6 | — | — | — | Option 2 | Chase 38770 |
| Rosaceae | Kerria japonica (L.) DC. | — | — | — | 260 | 8.4 | 1.8 | — | — | — | Option 2 | Chase 38777 |
| Rosaceae | Malus baccata (L). Borkh. var. jackii Borkh. | — | — | — | 75 | 6.7 | 1.9 | — | — | — | Option 2 | Chase 38773 |
| Rosaceae | Physocarpus opulifolius (L.) Maxim. | — | — | — | 163 | 8.3 | 2.2 | — | — | — | Option 2 | Chase 38776 |
| Rosaceae | Prunus prostrata Labill. | — | — | — | 155.6 | 7.1 | 1.7 | — | — | — | Option 2 | Chase 38785 |
| Rosaceae | Pyrus calleryana Decne. | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | All extractions failed | Chase 38791 |
| Rosaceae | Sanguisorba minor Scop. | — | — | — | 239.8 | 8.2 | 1.6 | — | — | — | Option 2 | Chase 38771 |
| Rosaceae | Sorbus koehneana C. K. Schneid. | — | — | — | 38.7 | 6.6 | 1.3 | — | — | — | Option 2 | Chase 38774 |
| Salviniaceae | Salvinia sp. Ség. | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | All extractions failed | Soltis and IEJ-T 3013 |
| Santalaceae | Phoradendron leucarpum (Raf.) Reveal & M. C. Johnst. | — | — | — | 37 | 4.1 | 0.3 | 59.8 | 2.3 | 0 | Option 2 | Soltis and Miles 2957 |
| Saxifragaceae | Astilbe chinensis (Maxim.) Franch. & Sav. | — | — | — | 60 | 7 | 1.6 | — | — | — | Option 2 | Chase 38784 |
| Saxifragaceae | Boykinia jamesii var. heucheriformis (Rydb.) Engl. | — | — | — | 63.9 | 6 | 2.1 | — | — | — | Option 2 | Chase 38782 |
| Saxifragaceae | Micranthes geum (L.) Small | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | All extractions failed | Chase 38790 |
| Saxifragaceae | Peltoboykinia watanabei (Yatabe) H. Hara | — | — | — | 70 | 7 | 1.7 | — | — | — | Option 2 | Chase 38787 |
| Saxifragaceae | Rodgersia podophylla A. Gray | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | All extractions failed | Chase 38783 |
| Saxifragaceae | Saxifraga geum L. var. gracilis | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | Tried but failed | All extractions failed | Chase 38790 |
| Saxifragaceae | Tellima breviflora Rydb. | — | — | — | 41.5 | 7.1 | 1.5 | — | — | — | Option 2 | Chase 38789 |
| Saxifragaceae | Tiarella polyphylla D. Don | — | — | — | 30 | 7.8 | 1.6 | — | — | — | Option 2 | Chase 38786 |
| Schisandraceae | Illicium floridanum J. Ellis | — | — | — | — | — | — | 25* | 4.8 | 0.8 | Option 3 | Soltis and Miles 2960 |
| Schlegeliaceae | Schlegelia parasitica (Sw.) Miers ex Griseb. | 18.2 | 8.2 | 1.5 | — | — | — | 7.8 | 2.1 | 0 | Option 1 | Soltis and Miles 2983 |
| Talinaceae | Talinum sp. Adans. | 8.6 | 7.3 | Missing | — | — | — | 15.4 | 2.3 | 0 | Option 1 | Soltis and Miles 2979 |
| Tetrachondraceae | Polypremum procumbens L. | 28.3 | 6.8 | 1.5 | — | — | — | 48.8 | 2.8 | 0.5 | Yes | Soltis and Miles 2989 |
Note: — = method was not tried on the species.
a
Gray cells are those with degraded RNA and either the library was not attempted or it failed. All samples were extracted from 60 to 100 mg of plant leaf tissue (for aquatics and succulents, 200 mg). The taxa that failed with all attempts were: Agrimonia eupatoria L. (Rosaceae), Cryptocarya alba (Molina) Looser (Lauraceae), Lachnanthes caroliniana (Lam.) Dandy (Haemodoraceae), Micranthes geum (L.) Small (Saxifragaceae), Opuntia pusilla (Haw.) Haw. (Cactaceae), Pyrus calleryana Decne. (Rosaceae), Rodgersia podophylla A. Gray (Saxifragaceae), Salvinia sp. Ség. (Salviniaceae), and Saxifraga geum L. var. gracilis (Saxifragaceae). Failed library construction despite having satisfactory metrics: Sprekelia formosissima (L.) Herb. (Amaryllidaceae) and Zephyranthes citrina Baker (Zephranthaceae).
b
RIN (RNA integrity number) is a measure from the Agilent 2100 Bioanalyzer that combines concentration and expected presence of subunits.
*
Marks the samples where an isolate was extracted from two to four extractions and pooled to be sequenced. (Not all extractions in Table 1 are in this table as not all were ran on the Bioanalyzer.)