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Fan, C., Deng, Q. & Zhu, T. F. Bioorthogonal information storage in l-DNA w\ ith a high-fidelity mirror-image Pfu DNA polymerase. Nat. Biotechnol. 39, 1548 1555 \(2021\).\r:2535)/F 4/H/N/Rect[240.716 349.206 245.921 336.66]/Subtype/Link/Type/Annot>> endobj 45 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 18. Liu, X. & Zhu, T. F. Sequencing mirror-image DNA chemically. Cell Chem. Biol.\ 25, 1151 1156 \(2018\).\r:2536)/F 4/H/N/Rect[247.157 349.206 252.362 336.66]/Subtype/Link/Type/Annot>> endobj 46 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 14. Fan, C., Deng, Q. & Zhu, T. F. Bioorthogonal information storage in l-DNA \ with a high-fidelity mirror-image Pfu DNA polymerase. Nat. Biotechnol. 39, 1548 1555 \(2021\).\r:2537)/F 4/H/N/Rect[276.289 317.706 281.494 305.16]/Subtype/Link/Type/Annot>> endobj 47 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 16. Jiang, W. et al. Mirror-image polymerase chain reaction. Cell Discov. 3, \n\ 17037 \(2017\).\r:2538)/F 4/H/N/Rect[284.313 317.706 289.518 305.16]/Subtype/Link/Type/Annot>> endobj 48 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 19. Muyzer, G. & Smalla, K. Application of denaturing gradient gel electrophoresis \ \(DGGE\) and temperature gradient gel electrophoresis \(TGGE\) in microbial ecology. Anton. Leeuw. 73, 127 141 \(1998\).\r:253\ 9)/F 4/H/N/Rect[196.801 223.206 202.006 210.66]/Subtype/Link/Type/Annot>> endobj 49 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd:Fig. 1 | Mirror-image selection of l-DNA aptamers targeting native human thromb\ in. a, Schematic overview of the direct, mirror-image selection scheme \(color\), which bypasses the need for chemically sy\ nthesizing mirror-image target mol:2540)/F 4/H/N/Rect[261.726 212.706 270.914 200.16]/Subtype/Link/Type/Annot>> endobj 50 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 15. Xu, W. et al. Total chemical synthesis of a thermostable enzyme capable of pol\ ymerase chain reaction. Cell Discov. 3, 17008 \(2017\).\r:2541)/F 4/H/N/Rect[67.2572 170.706 72.4622 158.16]/Subtype/Link/Type/Annot>> endobj 51 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 16. Jiang, W. et al. Mirror-image polymerase chain reaction. Cell Discov. 3, \ \n17037 \(2017\).\r:2542)/F 4/H/N/Rect[73.6983 170.706 78.9033 158.16]/Subtype/Link/Type/Annot>> endobj 52 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 20. Bock, L. C., Griffin, L. C., Latham, J. A., Vermaas, E. H. & Toole, J. J. Selec\ tion of single-stranded DNA molecules that bind and inhibit human thrombin. Nature 355, 564 566 \(1992\).\r:2543)/F 4/H/N/Rect[277.872 76.2059 283.077 63.6602]/Subtype/Link/Type/Annot>> endobj 53 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 21. Tasset, D. M., Kubik, M. F. & Steiner, W. Oligonucleotide inhibitors of human t\ hrombin that bind distinct epitopes. J. Mol. Biol. 272, 688 698 \(1997\).\r:2544)/F 4/H/N/Rect[284.313 76.2059 289.518 63.6602]/Subtype/Link/Type/Annot>> endobj 54 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 22. Dobbelstein, M. & Shenk, T. In vitro selection of RNA ligands for the ribosomal\ L22 protein associated with Epstein-Barr virus-expressed RNA \nby using randomized and cDNA-derived RNA libraries. J. 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Fan, C., Deng, Q. & Zhu, T. F. Bioorthogonal information storage in l-DN\ A with a high-fidelity mirror-image Pfu DNA polymerase. Nat. 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Liu, X. et al. RNA aptamers specific for bovine thrombin. J. Mol. 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Characterizing the optimized l-DNA aptamers from secondary selection.\ a,b, Secondary structures of the l-13 \(a\) and l-13t \(b, truncated version\) aptamers predicted by Mfold, with nu\ cleotides derived from the secondar:2576)/F 4/H/N/Rect[182.726 580.207 191.124 567.661]/Subtype/Link/Type/Annot>> endobj 130 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd:Fig. 4 | Characterizing the optimized l-DNA aptamers from secondary selection.\ a,b, Secondary structures of the l-13 \(a\) and l-13t \(b, truncated version\) aptamers predicted by Mfold, with n\ ucleotides derived from the seconda:2577)/F 4/H/N/Rect[127.754 538.207 136.943 525.661]/Subtype/Link/Type/Annot>> endobj 131 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd:Fig. 4 | Characterizing the optimized l-DNA aptamers from secondary selection.\ a,b, Secondary structures of the l-13 \(a\) and l-13t \(b, truncated version\) aptamers predicted by Mfold, with \ nucleotides derived from the second:2578)/F 4/H/N/Rect[171.602 517.821 180.976 503.778]/Subtype/Link/Type/Annot>> endobj 132 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd:Fig. 5 | l-DNA aptamer thrombin inhibitors. a, Schematic overview of inhibit\ ing native human thrombin enzymatic activity with the selected l-DNA aptamers, native human thrombin and a fluorogenic substra\ te, benzoyl-Phe-Val-Arg-AMC. b,:2579)/F 4/H/N/Rect[205.7 349.207 214.247 336.661]/Subtype/Link/Type/Annot>> endobj 133 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 29. Li, M.-L., Ren, Y.-J., Dong, M.-H. & Ren, W.-X. Design, synthesis and structura\ l exploration of novel fluorinated dabigatran derivatives as direct thrombin inhibitors. Eur. J. Med. Chem. 96, 122 138 \(20\ 15\).\r:2580)/F 4/H/N/Rect[219.218 338.707 224.423 326.161]/Subtype/Link/Type/Annot>> endobj 134 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd:Fig. 5 | l-DNA aptamer thrombin inhibitors. a, Schematic overview of inhibi\ ting native human thrombin enzymatic activity with the selected l-DNA aptamers, native human thrombin and a fluorogenic substr\ ate, benzoyl-Phe-Val-Arg-AMC. b:2581)/F 4/H/N/Rect[206.256 307.821 215.444 293.778]/Subtype/Link/Type/Annot>> endobj 135 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd:Fig. 5 | l-DNA aptamer thrombin inhibitors. a, Schematic overview of inhib\ iting native human thrombin enzymatic activity with the selected l-DNA aptamers, native human thrombin and a fluorogenic subst\ rate, benzoyl-Phe-Val-Arg-AMC. b:2582)/F 4/H/N/Rect[166.372 234.321 175.56 220.278]/Subtype/Link/Type/Annot>> endobj 136 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd:Fig. 5 | l-DNA aptamer thrombin inhibitors. a, Schematic overview of inhi\ biting native human thrombin enzymatic activity with the selected l-DNA aptamers, native human thrombin and a fluorogenic subs\ trate, benzoyl-Phe-Val-Arg-AMC. :2583)/F 4/H/N/Rect[174.185 118.207 182.582 105.661]/Subtype/Link/Type/Annot>> endobj 137 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd:Fig. 5 | l-DNA aptamer thrombin inhibitors. a, Schematic overview of inh\ ibiting native human thrombin enzymatic activity with the selected l-DNA aptamers, native human thrombin and a fluorogenic sub\ strate, benzoyl-Phe-Val-Arg-AMC. :2584)/F 4/H/N/Rect[276.925 76.2068 286.3 63.6611]/Subtype/Link/Type/Annot>> endobj 138 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 30. Khanin, M. A., Rakov, D. V. & Kogan, A. E. Mathematical model for the blood coa\ gulation prothrombin time test. Thromb. Res. 89, 227 232 \(1998\).\r:2585)/F 4/H/N/Rect[134.959 55.2068 140.164 42.6611]/Subtype/Link/Type/Annot>> endobj 139 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 31. Nutiu, R. & Li, Y. F. Structure-switching signaling aptamers. J. Am. Chem. 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Li, L.-L., Ge, P., Selvin, P. R. & Lu, Y. Direct detection of adenosine in undi\ luted serum using a luminescent aptamer sensor attached to a terbium complex. Anal. Chem. 84, 7852 7856 \(2012\).\r:2589)/F 4/H/N/Rect[105.352 265.207 110.557 252.661]/Subtype/Link/Type/Annot>> endobj 167 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd:Fig. 6 | l-DNA aptamer sensor and western blot. a, Schematic overview of d\ etecting native human thrombin with the l-DNA aptamer sensor, by hybridizing the l-9-1t aptamer strand \(cyan\) with an l-D\ NA fluorophore strand labeled with:2590)/F 4/H/N/Rect[187.427 233.707 196.615 221.161]/Subtype/Link/Type/Annot>> endobj 168 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 33. Barra, G. B. et al. EDTA-mediated inhibition of DNases protects circulating cel\ l-free DNA from ex vivo degradation in blood samples. Clin. Biochem. 48, 976 981 \(2015\).\r:2591)/F 4/H/N/Rect[66.0573 170.707 71.2622 158.161]/Subtype/Link/Type/Annot>> endobj 169 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd:Fig. 6 | l-DNA aptamer sensor and western blot. a, Schematic overview of \ detecting native human thrombin with the l-DNA aptamer sensor, by hybridizing the l-9-1t aptamer strand \(cyan\) with an l-\ DNA fluorophore strand labeled wit:2592)/F 4/H/N/Rect[155.095 139.207 163.493 126.661]/Subtype/Link/Type/Annot>> endobj 170 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 6. Williams, K. P. et al. Bioactive and nuclease-resistant l-DNA ligand of vas\ opressin. Proc. Natl. Acad. Sci. USA 94, 11285 11290 \(1997\).\r:2593)/F 4/H/N/Rect[327.175 223.207 329.778 210.661]/Subtype/Link/Type/Annot>> endobj 171 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 34. Kratschmer, C. & Levy, M. Effect of chemical modifications on aptamer stability\ in serum. Nucleic Acid Ther. 27, 335 344 \(2017\).\r:2594)/F 4/H/N/Rect[331.014 223.207 336.219 210.661]/Subtype/Link/Type/Annot>> endobj 172 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd:Fig. 6 | l-DNA aptamer sensor and western blot. a, Schematic overview of\ detecting native human thrombin with the l-DNA aptamer sensor, by hybridizing the l-9-1t aptamer strand \(cyan\) with an l\ -DNA fluorophore strand labeled wi:2595)/F 4/H/N/Rect[399.807 170.707 409.181 158.161]/Subtype/Link/Type/Annot>> endobj 173 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd:Fig. 6 | l-DNA aptamer sensor and western blot. a, Schematic overview o\ f detecting native human thrombin with the l-DNA aptamer sensor, by hybridizing the l-9-1t aptamer strand \(cyan\) with an l\ -DNA fluorophore strand labeled w:2596)/F 4/H/N/Rect[418.687 107.707 427.103 95.1611]/Subtype/Link/Type/Annot>> endobj 174 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd:Fig. 6 | l-DNA aptamer sensor and western blot. a, Schematic overview \ of detecting native human thrombin with the l-DNA aptamer sensor, by hybridizing the l-9-1t aptamer strand \(cyan\) with an \ l-DNA fluorophore strand labeled :2597)/F 4/H/N/Rect[303.638 86.7068 311.58 74.1611]/Subtype/Link/Type/Annot>> endobj 175 0 obj <>/BS<>/Border[0 0 0]/F 4/H/N/Rect[244.301 33.864 352.897 25.1497]/Subtype/Link/Type/Annot>> endobj 176 0 obj <>/ExtGState<>/Font<>/ProcSet[/PDF/Text/ImageB/ImageC]/Properties<>/Shading<>/XObject<>>> endobj 177 0 obj <>stream x}is$q?x3N#BKQ!R ,0cUw/3{+%0ʻ?}z}wWz~sۣO> ><;b1/VQ-Z2?A_~Sy%5iW_}]sڮr޸w'k^s?kC3\ᗶ[t^mB[oꂠݬX]_m-%k|zdǿE"V.9`Z' 9 ~vpkO+\!p ҟ8dՆ\H9Luv'Zmu )]ޮoU|T9 ;HQΪw+{zV!nUd#Pa(eEdl\>W~fD7k -JҍH Hj@l{wӛ]C%䢣 hC NvDU2%K!O8t[٪f̌x-q- WQ|dU+.k „VJpĝƙIi*}ͬ0o%z1yrohy]o/uO<ˋOFV~7ag q񰕓VE֒@njCg.Ba| dZoK?9;k7"PkJ'bT$a{hٶѮAτs9*4*_8S$2SbyP˛ץr^k\ lo^Q7v4ockrɟ3ryHsap^Rel¹p v͍Q uJ0IUP!Yh Y>~?WշOv ye$_oW_}i@Z}͊&E5bU'ߤ4r7N{X]:\;)ps{j57+Im461џ,p4F^2:$Ue7l5m@!$6lQE5Uֿ8x. `lxjuxcꊻt^F]0M-:)-><.[cp-vౝB/@{'V wX:vf5eLdm$m GmjyFA@BbHd]C.ˤVDT *N/ "Ѫ/a˷.Զqu= R};mZ,%tGʛV:'6br8z^h=JlES* :xZin^al^f%to.3a+SfA^ҷ^Q `lRtM_rNBNS<8|XXCXwK 2zqM104Jq'煮v8MQH/J^OGWLPqLiD89́5{\*[(vb6'A@gv"u:ϯ0 (&t`&Yn Vu%f)Kȥ[+5:h[d7E,G6׃qFtR-d5.$Ɛo(wt')= w虾a^Vg Jݸ1Mk5 :VQe`Ci0zj&0?I&' bZ$[Jo k[%% wâpk2G5SO@@~'XB& JHR$a+:uNHz[%hBY}aNqGjbgȉYVRCƶ& W0ʓudK #kB[xWȜ_ ǶNW[-;F=2>%1-LTr0u>fU}AM*>$Mxt4WJ( Mֳpy4MDuS$~Lޔ_-"sdS11f@Gfڅ1׹3] v ̐fWTwm8Sץ͏u,gWpoWUԙg EtR]wSB"W'+q9 `vT#[fO_I gua&@է wKzN]p04m7wO50_HMVK:A.ݤ/, 5Qd7YYAp7ϿXG^Zj4Ȃ[+ !1 DP^n~wt ?t@u{qߧ02l0 SN(f6N>9,!H΁#t؊ŬIҵq_y^{鄗`PHJ`]ʚ 흳F#,6rФ1 fa3chNY FlDD}ATFڗ]akNT5ѺA'3A$N $":AN~"}}c~u9Q`+nk%`_acˣ0I핯j$g'QЕF^S@4R̴x J )K&18 5:U(GŲMDP]ÏLت,<#/`!9 cЁPdVdi6rO@ǨKm4 Dw!ڶֆ?iC#D%aa2I H Td#`Iq\~R,2 "y>mjo1R(9)TF_z⽅?(#7465!tLXC$jFƱ#DHDD:i6X3MVĢW?Ýa`y*,d$Ɏ9ř&{ :2d47YuA KkFALsUK* R1&hch(QXZ<$밋 "- Xp&P2*{sh"gT4pli Jg@aJ9<xPM2ڊ8e&C%A} b7Z˨!4Pr{QcGs99d('-&fDl6dǣ`踑4y vh[;( !!EOdƤtؙ@ m\AVϬbaǂ`O`{*52> Ú pAZO#A79!n`eP7`$"t!s 2` I H* [Ă0DRŘ8d8DZj@V C^!pGW$B5oai @ ՇIlc̓ F!1/htyC@uqBPD ϭr9B_aJ0`Z'GI[ðA0.{2,T/%*b1= !H R !p.[f"1_3M4.%(dz#THaON7BJXRCNХW :F?,0aFPBêd 2%phLZҰ4Gqq xM.W 3Iq6W%E͡W4 pϜ"|}~p6F(tCN>g8>=Zv B9M5bYvr(#aRZBG_DEY3CG Cvy>f1|$rg1?R)|%| |t͜1pFL)VNCg)6բ$DB 0U_I6Z9ù::|l It(-']EVMa-`#@*BNY ̢[R8C,A^DآxL0'M,ęX6c|H0,5K0'EHXnͣZ#B^;p j<$A @l<[fa`!OtcCvRB0mA e+pQ Ո *JFAeU$TBuIbKB]HQR*X|2X`b N\*i},t*fLm1sp Ztkdڇ#סL㛴.xyJv -E躓.psk:Hsd>] H0@*~gti*ɞ R"N. 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Fang, X. H. & Tan, W. H. Aptamers generated from cell-SELEX for molecular medic\ ine: a chemical biology approach. Acc. Chem. Res. 43, 48 57 \(2010\).\r:2599)/F 4/H/N/Rect[279.992 527.707 285.197 515.161]/Subtype/Link/Type/Annot>> endobj 200 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 36. Li, S. et al. Identification of an aptamer targeting hnRNP A1 by tissue slide-b\ ased SELEX. J. Pathol. 218, 327 336 \(2009\).\r:2600)/F 4/H/N/Rect[286.433 527.707 291.638 515.161]/Subtype/Link/Type/Annot>> endobj 201 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 9. Sczepanski, J. T. & Joyce, G. F. Binding of a structured d-RNA molecule by an\ l-RNA aptamer. J. Am. Chem. Soc. 135, 13290 13293 \(2013\).\r:2601)/F 4/H/N/Rect[167.313 401.707 169.915 389.161]/Subtype/Link/Type/Annot>> endobj 202 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 10. Umar, M. I. & Kwok, C. K. 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Che\ m 5, 848 857 \(2019\).\r:2605)/F 4/H/N/Rect[210.097 380.707 215.302 368.161]/Subtype/Link/Type/Annot>> endobj 206 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 39. Wang, Z., Xu, W., Liu, L. & Zhu, T. F. A synthetic molecular system capable of \ mirror-image genetic replication and transcription. Nat. Chem. 8, \n698 704 \(2016\).\r:2606)/F 4/H/N/Rect[148.928 338.707 154.133 326.161]/Subtype/Link/Type/Annot>> endobj 207 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 43. Chen, J., Chen, M. & Zhu, T. F. Translating protein enzymes without aminoacyl-t\ RNA synthetases. Chem 7, 786 798 \(2021\).\r:2607)/F 4/H/N/Rect[156.952 338.707 162.157 326.161]/Subtype/Link/Type/Annot>> endobj 208 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 44. Mattheakis, L. C., Bhatt, R. R. & Dower, W. J. An in vitro polysome display sys\ tem for identifying ligands from very large peptide libraries. Proc. Natl. Acad. Sci. 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Caton-Williams, J., Hoxhaj, R., Fiaz, B. & Huang, Z. Use of a novel 5 2-regiosel\ ective phosphitylating reagent for one-pot synthesis of nucleoside 5 2-triphosphates from unprotected nucleosides. Curr. Protoc.\ Nucleic Acid Chem. 52, 1.30.3:2618)/F 4/H/N/Rect[161.617 649.422 165.815 638.097]/Subtype/Link/Type/Annot>> endobj 242 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 15. Xu, W. et al. Total chemical synthesis of a thermostable enzyme capable of po\ lymerase chain reaction. Cell Discov. 3, 17008 \(2017\).\r:2619)/F 4/H/N/Rect[80.9639 622.827 85.1614 612.709]/Subtype/Link/Type/Annot>> endobj 243 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 15. Xu, W. et al. Total chemical synthesis of a thermostable enzyme capable of p\ olymerase chain reaction. Cell Discov. 3, 17008 \(2017\).\r:2620)/F 4/H/N/Rect[69.7666 614.127 73.9642 604.009]/Subtype/Link/Type/Annot>> endobj 244 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 16. Jiang, W. et al. Mirror-image polymerase chain reaction. Cell Discov. 3,\ \n17037 \(2017\).\r:2621)/F 4/H/N/Rect[74.9611 614.127 79.1586 604.009]/Subtype/Link/Type/Annot>> endobj 245 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 52. Pollard, J., Bell, S. D. & Ellington, A. D. Design, synthesis, and amplificatio\ n of DNA pools for in vitro selection. Curr. Protoc. Nucleic Acid Chem. 9, 9.2.1 9.2.23 \(2000\).\r:2622)/F 4/H/N/Rect[121.635 431.427 125.833 421.309]/Subtype/Link/Type/Annot>> endobj 246 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 53. Chizzolini, F. et al. Large phenotypic enhancement of structured random RNA poo\ ls. J. Am. Chem. Soc. 142, 1941 1951 \(2020\).\r:2623)/F 4/H/N/Rect[201.779 422.727 205.976 412.609]/Subtype/Link/Type/Annot>> endobj 247 0 obj <>/BS<>/Border[0 0 0]/Dest(SpringerNature_NatBio_1337.indd: 54. Golden, B. 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