SUPPLEMENTARY MATERIAL111Equation Chapter 1 Section 1

Computational selection of RNA aptamer against angiopoietin-2 and

experimental evaluation

Wen-Pin Hua,b*, Jangam Vikram Kumara*, Chun-Jen Huangc, Wen-Yih Chend

a Department of Biomedical Informatics, Asia University, Taichung City, Taiwan b Department of Medical Laboratory Science and Biotechnology, China Medical University, Taichung City, Taiwan

c Graduate Institute of Biomedical Engineering, National Central University, Jhong-Li City, Taiwan

d Department of Chemical and Materials Engineering, National Central University, Jhong-Li City, Taiwan

Corresponding author: Wen-Pin Hu; Jangam Vikram Kumar

Department of Biomedical Informatics, Asia University

500, Lioufeng Rd., Wufeng, Taichung 41354, Taiwan

TEL: +886-4-23323456 ext.20022; FAX: +886-4-23305737

E-mail: wenpinhu@asia.edu.tw (W.P. Hu); vikramkumar.bioinf@gmail.com (J.V.

Kumar)

1. Materials and methods1.1 Computational assay

Figure S1.Model of the 3D Structure of Seq15. This model is generated by using the CentroidFold and RNAComposer webservers.

1.2 The library of mutated RNA sequences

Table S1.Nucleotides of three RNA aptamers involved in the binding interface with Ang2. Nucleotides at position 10 to 16 and 33 to 99 were marked with bold, underlined text.

Name Nucleotides involved in binding interface with Ang2

Seq1 A4 G5 C6 C7 U8 C9 A10 A13 G14 C15 U16 C17 A18 G30 C31 C32 U33 G34 G35 A36

Seq2 U2 A3 A4 C5 C6 A7 U8 C9 A10 G11 C12 U13 C14 A15 U16A39 C40

Seq3 C28 U29 C30 A31 C32 C33 C34 A35 C36 A37 C38 C39 G40

Seq4 C1 C2 A3 C4 C5 G6 A7 U8 C9 G10 C11 A12 U13 C14 A15 G16

Seq5 U9 C10 U11 G18 C19 C20 G21 A22 U23 C24 A25 U26 C27 G29 C30 G31 C32 U33

Seq6 A5 C6 C7 A8 C9 G10 C11 U14 A15 U16 C17 A18 G19 C20 U21 A22 A23 C34 U35 A36

Seq7 C6 C7 A8 G9 U10 C11 A12 C13 C14 A15 U16

Seq8 C4 C5 A6 A7 G8 C10 U11 C12 A13 C14 G15 U16 U17

Seq9 G1 G2 A3 G4 C5 G6 C7 A8 A9 G25 A26 A27 C28 U29 C30 C31 G32

Seq10 C13 U14 U15 A16 G29 A30 A31 C32 U33 C34 C35 A36 G37 A38

Table S1. (Continued) Nucleotides of three RNA aptamers involved in binding interface with Ang2. Nucleotides at position 10 to 16 and 33 to 99 were marked with bold, underlined text.Seq11 A26 A27 C28 U29 C30 C31 A32 U33 G34 C35 A36

Seq12 G23 U24 U25 G26 G28 U29 C30 U31 C32 G33 U34 C35 G36 A37 A38

Seq13 C1 A2 C3 U4 C5 A6 G7 C8 G9 C10 C11 C12 U13 G14 C15 G16 A17 A18 C32 C39 U40

Seq14 G22 A23 A24 C25 U26 C27 C28 U29 G30 C32 C33 C34 U35 C36 U37 A38 C39

Seq15 G10 C11 G12 G13 A14 C20 U21 C22 A23 U24 C37 C38

Seq16 C13 C14 A15 C16 U17 C20 C21 A22 A23 C24 C25 U27

1.3 SPR imaging apparatus

The SPR platform is a self- referencing SPR imaging sensor with polarization

contrast, which is able to detect the smallest signal corresponding to the change in the

refractive index unit (RIU) better than10-6 within the operation range of 0.011 RIU.

The p-polarized narrow-band light beam (central emission wavelength is 750 nm)

impinges on a glass substrate and excites SPs at the metal-dielectric interface. It also

can be illustrated in terms of protein surface coverage that the sensor can detect the

changes that are as small as 0.2 pg/mm2. The SPRi can be utilized by conducting

traditional measurements or performing a high-throughput analysis with the use of an

array-formatted chip. The SPR chips were prepared by using clean BK7 glass

substrates. The glass substrates were pre-coated with an adhesion layer of chromium

(thickness approx. 2 nm), and an active gold layer with a thickness of 48 nm was

subsequently coated via an evaporation deposition process at pressures below 110-6

Torr. All experiments were performed at a constant temperature of 25°C and at a flow

rate of 50 μl/min. Six flow channels were used in the SPR experiments, and one

channel was used as a reference channel.

2. Results and discussion2.1. Selection of aptamers from computational results

Table S2.The simulation results of 189 mutant RNA aptamer sequences. The mutated positions in the sequences and the ZRNAK scores of three selected aptamers were marked with bold letters.

Type Sequence(5-3) ZRANK Score

Mut

ant s

eque

nce

of S

eq1

ACUAGCCUCUUCAGCUCAUGUGCCCCUCCGCCAGGAUCAC -66.174

ACUAGCCUCUUCAGCUCAUGUGCCCCUCCGCCGGGAUCAC -73.649

ACUAGCCUCUUCAGCUCAUGUGCCCCUCCGCCCGGAUCAC -69.671

ACUAGCCUCGUCAGCUCAUGUGCCCCUCCGCCAGGAUCAC -63.625

ACUAGCCUCGUCAGCUCAUGUGCCCCUCCGCCGGGAUCAC -61.696

ACUAGCCUCGUCAGCUCAUGUGCCCCUCCGCCCGGAUCAC -71.018

ACUAGCCUCCUCAGCUCAUGUGCCCCUCCGCCAGGAUCAC -74.2

ACUAGCCUCCUCAGCUCAUGUGCCCCUCCGCCGGGAUCAC -62.316

ACUAGCCUCCUCAGCUCAUGUGCCCCUCCGCCCGGAUCAC -66.705

ACUAGCCUCAACAGCUCAUGUGCCCCUCCGCCUAGAUCAC -73.514

ACUAGCCUCAACAGCUCAUGUGCCCCUCCGCCUCGAUCAC -74.701

ACUAGCCUCAACAGCUCAUGUGCCCCUCCGCCUUGAUCAC -87.666

ACUAGCCUCAGCAGCUCAUGUGCCCCUCCGCCUAGAUCAC -66.493

ACUAGCCUCAGCAGCUCAUGUGCCCCUCCGCCUCGAUCAC -66.839

ACUAGCCUCAGCAGCUCAUGUGCCCCUCCGCCUUGAUCAC -61.193

ACUAGCCUCACCAGCUCAUGUGCCCCUCCGCCUAGAUCAC -87.829

ACUAGCCUCACCAGCUCAUGUGCCCCUCCGCCUCGAUCAC -74.337

ACUAGCCUCACCAGCUCAUGUGCCCCUCCGCCUUGAUCAC -65.241

ACUAGCCUCAUAAGCUCAUGUGCCCCUCCGCCUGAAUCAC -71.17

ACUAGCCUCAUAAGCUCAUGUGCCCCUCCGCCUGCAUCAC -76.63

ACUAGCCUCAUAAGCUCAUGUGCCCCUCCGCCUGUAUCAC -68.573

ACUAGCCUCAUGAGCUCAUGUGCCCCUCCGCCUGAAUCAC -74.78

ACUAGCCUCAUGAGCUCAUGUGCCCCUCCGCCUGCAUCAC -72.671

ACUAGCCUCAUGAGCUCAUGUGCCCCUCCGCCUGUAUCAC -73.072

ACUAGCCUCAUUAGCUCAUGUGCCCCUCCGCCUGAAUCAC -68.47

ACUAGCCUCAUUAGCUCAUGUGCCCCUCCGCCUGCAUCAC -83.747

ACUAGCCUCAUUAGCUCAUGUGCCCCUCCGCCUGUAUCAC -64.297

ACUAGCCUCAUCUGCUCAUGUGCCCCUCCGCCUGGCUCAC -77.279

ACUAGCCUCAUCUGCUCAUGUGCCCCUCCGCCUGGGUCAC -70.501

ACUAGCCUCAUCUGCUCAUGUGCCCCUCCGCCUGGUUCAC -73.393

Table S2.(Continued) The simulation results of 189 mutant RNA aptamer sequences. The mutated positions in the sequences and the ZRNAK scores of three selected aptamers were marked with bold letters.

Type Sequence(5-3) ZRANK Score

Mut

ant s

eque

nce

of S

eq1

ACUAGCCUCAUCGGCUCAUGUGCCCCUCCGCCUGGCUCAC -69.012

ACUAGCCUCAUCGGCUCAUGUGCCCCUCCGCCUGGGUCAC -60.148

ACUAGCCUCAUCGGCUCAUGUGCCCCUCCGCCUGGUUCAC -71.464

ACUAGCCUCAUCCGCUCAUGUGCCCCUCCGCCUGGCUCAC -79.954

ACUAGCCUCAUCCGCUCAUGUGCCCCUCCGCCUGGGUCAC -76.995

ACUAGCCUCAUCCGCUCAUGUGCCCCUCCGCCUGGUUCAC -78.571

ACUAGCCUCAUCAACUCAUGUGCCCCUCCGCCUGGAACAC -77.097

ACUAGCCUCAUCAACUCAUGUGCCCCUCCGCCUGGACCAC -67.97

ACUAGCCUCAUCAACUCAUGUGCCCCUCCGCCUGGAGCAC -74.329

ACUAGCCUCAUCACCUCAUGUGCCCCUCCGCCUGGAACAC -65.604

ACUAGCCUCAUCACCUCAUGUGCCCCUCCGCCUGGACCAC -70.131

ACUAGCCUCAUCACCUCAUGUGCCCCUCCGCCUGGAGCAC -73.143

ACUAGCCUCAUCAUCUCAUGUGCCCCUCCGCCUGGAACAC -72.094

ACUAGCCUCAUCAUCUCAUGUGCCCCUCCGCCUGGACCAC -66.34

ACUAGCCUCAUCAUCUCAUGUGCCCCUCCGCCUGGAGCAC -78.013

ACUAGCCUCAUCAGAUCAUGUGCCCCUCCGCCUGGAUAAC -69.839

ACUAGCCUCAUCAGAUCAUGUGCCCCUCCGCCUGGAUGAC -67.988

ACUAGCCUCAUCAGAUCAUGUGCCCCUCCGCCUGGAUUAC -78.518

ACUAGCCUCAUCAGGUCAUGUGCCCCUCCGCCUGGAUAAC -66.59

ACUAGCCUCAUCAGGUCAUGUGCCCCUCCGCCUGGAUGAC -68.432

ACUAGCCUCAUCAGGUCAUGUGCCCCUCCGCCUGGAUUAC -77.178

ACUAGCCUCAUCAGUUCAUGUGCCCCUCCGCCUGGAUAAC -66.492

ACUAGCCUCAUCAGUUCAUGUGCCCCUCCGCCUGGAUGAC -61.954

ACUAGCCUCAUCAGUUCAUGUGCCCCUCCGCCUGGAUUAC -66.717

ACUAGCCUCAUCAGCACAUGUGCCCCUCCGCCUGGAUCCC -84.072

ACUAGCCUCAUCAGCACAUGUGCCCCUCCGCCUGGAUCGC -67.44

ACUAGCCUCAUCAGCACAUGUGCCCCUCCGCCUGGAUCUC -67.844

ACUAGCCUCAUCAGCGCAUGUGCCCCUCCGCCUGGAUCCC -68.77

ACUAGCCUCAUCAGCGCAUGUGCCCCUCCGCCUGGAUCGC -64.424

ACUAGCCUCAUCAGCGCAUGUGCCCCUCCGCCUGGAUCUC -65.86

ACUAGCCUCAUCAGCCCAUGUGCCCCUCCGCCUGGAUCCC -65.771

ACUAGCCUCAUCAGCCCAUGUGCCCCUCCGCCUGGAUCGC -77.968

ACUAGCCUCAUCAGCCCAUGUGCCCCUCCGCCUGGAUCUC -74.137

Table S2.(Continued) The simulation results of 189 mutant RNA aptamer sequences. The mutated positions in the sequences and the ZRNAK scores of three selected aptamers were marked with bold letters.

Type Sequence(5-3) ZRANK Score

Mut

ant s

eque

nce

of S

eq2

UUAACCAUCUGCUCAUGGCCCCUGCCCUCUCAUGGACCAC -87.021

UUAACCAUCUGCUCAUGGCCCCUGCCCUCUCAGGGACCAC -78.846

UUAACCAUCUGCUCAUGGCCCCUGCCCUCUCACGGACCAC -73.147

UUAACCAUCGGCUCAUGGCCCCUGCCCUCUCAUGGACCAC -61.103

UUAACCAUCGGCUCAUGGCCCCUGCCCUCUCAGGGACCAC -71.789

UUAACCAUCGGCUCAUGGCCCCUGCCCUCUCACGGACCAC -73.519

UUAACCAUCCGCUCAUGGCCCCUGCCCUCUCAUGGACCAC -68.534

UUAACCAUCCGCUCAUGGCCCCUGCCCUCUCAGGGACCAC -67.404

UUAACCAUCCGCUCAUGGCCCCUGCCCUCUCACGGACCAC -71.113

UUAACCAUCAACUCAUGGCCCCUGCCCUCUCAAAGACCAC -83.898

UUAACCAUCAACUCAUGGCCCCUGCCCUCUCAAUGACCAC -70.853

UUAACCAUCAACUCAUGGCCCCUGCCCUCUCAACGACCAC -68.273

UUAACCAUCAUCUCAUGGCCCCUGCCCUCUCAAAGACCAC -87.682

UUAACCAUCAUCUCAUGGCCCCUGCCCUCUCAAUGACCAC -63.614

UUAACCAUCAUCUCAUGGCCCCUGCCCUCUCAACGACCAC -66.266

UUAACCAUCACCUCAUGGCCCCUGCCCUCUCAAAGACCAC -72.281

UUAACCAUCACCUCAUGGCCCCUGCCCUCUCAAUGACCAC -76.843

UUAACCAUCACCUCAUGGCCCCUGCCCUCUCAACGACCAC -77.119

UUAACCAUCAGAUCAUGGCCCCUGCCCUCUCAAGAACCAC -82.028

UUAACCAUCAGAUCAUGGCCCCUGCCCUCUCAAGUACCAC -58.122

UUAACCAUCAGAUCAUGGCCCCUGCCCUCUCAAGCACCAC -97.609

UUAACCAUCAGUUCAUGGCCCCUGCCCUCUCAAGAACCAC -69.531

UUAACCAUCAGUUCAUGGCCCCUGCCCUCUCAAGUACCAC -70.08

UUAACCAUCAGUUCAUGGCCCCUGCCCUCUCAAGCACCAC -78.163

UUAACCAUCAGGUCAUGGCCCCUGCCCUCUCAAGAACCAC -87.09

UUAACCAUCAGGUCAUGGCCCCUGCCCUCUCAAGUACCAC -84.423

UUAACCAUCAGGUCAUGGCCCCUGCCCUCUCAAGCACCAC -67.203

UUAACCAUCAGCACAUGGCCCCUGCCCUCUCAAGGUCCAC -69.167

UUAACCAUCAGCACAUGGCCCCUGCCCUCUCAAGGGCCAC -70.366

UUAACCAUCAGCACAUGGCCCCUGCCCUCUCAAGGCCCAC -73.611

UUAACCAUCAGCGCAUGGCCCCUGCCCUCUCAAGGUCCAC -74.263

UUAACCAUCAGCGCAUGGCCCCUGCCCUCUCAAGGGCCAC -79.564

Table S2.(Continued) The simulation results of 189 mutant RNA aptamer sequences. The mutated positions in the sequences and the ZRNAK scores of three selected aptamers were marked with bold letters.

Type Sequence(5-3) ZRANK Score

Mut

ant s

eque

nce

of S

eq2

UUAACCAUCAGCGCAUGGCCCCUGCCCUCUCAAGGCCCAC -79.122

UUAACCAUCAGCCCAUGGCCCCUGCCCUCUCAAGGUCCAC -69.499

UUAACCAUCAGCCCAUGGCCCCUGCCCUCUCAAGGGCCAC -74.227

UUAACCAUCAGCCCAUGGCCCCUGCCCUCUCAAGGCCCAC -68.816

UUAACCAUCAGCUAAUGGCCCCUGCCCUCUCAAGGAACAC -68.767

UUAACCAUCAGCUAAUGGCCCCUGCCCUCUCAAGGAUCAC -77.011

UUAACCAUCAGCUAAUGGCCCCUGCCCUCUCAAGGAGCAC -64.889

UUAACCAUCAGCUUAUGGCCCCUGCCCUCUCAAGGAACAC -79.081

UUAACCAUCAGCUUAUGGCCCCUGCCCUCUCAAGGAUCAC -84.05

UUAACCAUCAGCUUAUGGCCCCUGCCCUCUCAAGGAGCAC -71.727

UUAACCAUCAGCUGAUGGCCCCUGCCCUCUCAAGGAACAC -77.511

UUAACCAUCAGCUGAUGGCCCCUGCCCUCUCAAGGAUCAC -67.982

UUAACCAUCAGCUGAUGGCCCCUGCCCUCUCAAGGAGCAC -72.09

UUAACCAUCAGCUCUUGGCCCCUGCCCUCUCAAGGACUAC -69.207

UUAACCAUCAGCUCUUGGCCCCUGCCCUCUCAAGGACGAC -72.46

UUAACCAUCAGCUCGUGGCCCCUGCCCUCUCAAGGACAAC -70.948

UUAACCAUCAGCUCGUGGCCCCUGCCCUCUCAAGGACUAC -72.441

UUAACCAUCAGCUCGUGGCCCCUGCCCUCUCAAGGACGAC -67.28

UUAACCAUCAGCUCCUGGCCCCUGCCCUCUCAAGGACAAC -67.681

UUAACCAUCAGCUCCUGGCCCCUGCCCUCUCAAGGACUAC -66.611

UUAACCAUCAGCUCCUGGCCCCUGCCCUCUCAAGGACGAC -78.931

UUAACCAUCAGCUCAAGGCCCCUGCCCUCUCAAGGACCUC -68.63

UUAACCAUCAGCUCAAGGCCCCUGCCCUCUCAAGGACCGC -63.629

UUAACCAUCAGCUCAAGGCCCCUGCCCUCUCAAGGACCCC -66.074

UUAACCAUCAGCUCAGGGCCCCUGCCCUCUCAAGGACCUC -86.223

UUAACCAUCAGCUCAGGGCCCCUGCCCUCUCAAGGACCGC -73.089

UUAACCAUCAGCUCAGGGCCCCUGCCCUCUCAAGGACCCC -67.327

UUAACCAUCAGCUCACGGCCCCUGCCCUCUCAAGGACCUC -66.499

UUAACCAUCAGCUCACGGCCCCUGCCCUCUCAAGGACCGC -72.857

UUAACCAUCAGCUCACGGCCCCUGCCCUCUCAAGGACCCC -60.727

Table S2.(Continued) The simulation results of 189 mutant RNA aptamer sequences. The mutated positions in the sequences and the ZRNAK scores of three selected aptamers were marked with bold letters.

Type Sequence(5-3) ZRANK Score

Mut

ant s

eque

nce

of S

eq15

GAGGACGAUACGGACUAGCCUCAUCAGCUCAUAUGCCCCUC -72.04

GAGGACGAUACGGACUAGCCUCAUCAGCUCAUUUGCCCCUC -71.647

GAGGACGAUACGGACUAGCCUCAUCAGCUCAUCUGCCCCUC -78.706

GAGGACGAUUCGGACUAGCCUCAUCAGCUCAUAUGCCCCUC -68.452

GAGGACGAUUCGGACUAGCCUCAUCAGCUCAUUUGCCCCUC -88.897

GAGGACGAUUCGGACUAGCCUCAUCAGCUCAUCUGCCCCUC -71.47

GAGGACGAUCCGGACUAGCCUCAUCAGCUCAUAUGCCCCUC -63.921

GAGGACGAUCCGGACUAGCCUCAUCAGCUCAUUUGCCCCUC -62.649

GAGGACGAUCCGGACUAGCCUCAUCAGCUCAUCUGCCCCUC -76.687

GAGGACGAUGAGGACUAGCCUCAUCAGCUCAUGAGCCCCUC -88.326

GAGGACGAUGAGGACUAGCCUCAUCAGCUCAUGGGCCCCUC -79.479

GAGGACGAUGAGGACUAGCCUCAUCAGCUCAUGCGCCCCUC -76.345

GAGGACGAUGGGGACUAGCCUCAUCAGCUCAUGAGCCCCUC -86.981

GAGGACGAUGGGGACUAGCCUCAUCAGCUCAUGGGCCCCUC -76.127

GAGGACGAUGGGGACUAGCCUCAUCAGCUCAUGCGCCCCUC -76.427

GAGGACGAUGUGGACUAGCCUCAUCAGCUCAUGAGCCCCUC -75.317

GAGGACGAUGUGGACUAGCCUCAUCAGCUCAUGGGCCCCUC -76.279

GAGGACGAUGUGGACUAGCCUCAUCAGCUCAUGCGCCCCUC -73.258

GAGGACGAUGCAGACUAGCCUCAUCAGCUCAUGUACCCCUC -78.506

GAGGACGAUGCAGACUAGCCUCAUCAGCUCAUGUUCCCCUC -81.902

GAGGACGAUGCAGACUAGCCUCAUCAGCUCAUGUCCCCCUC -66.779

GAGGACGAUGCUGACUAGCCUCAUCAGCUCAUGUACCCCUC -86.916

GAGGACGAUGCUGACUAGCCUCAUCAGCUCAUGUUCCCCUC -89.858

GAGGACGAUGCUGACUAGCCUCAUCAGCUCAUGUCCCCCUC -76.02

GAGGACGAUGCCGACUAGCCUCAUCAGCUCAUGUACCCCUC -70.982

GAGGACGAUGCCGACUAGCCUCAUCAGCUCAUGUUCCCCUC -72.649

GAGGACGAUGCCGACUAGCCUCAUCAGCUCAUGUCCCCCUC -93.335

GAGGACGAUGCGAACUAGCCUCAUCAGCUCAUGUGACCCUC -81.726

GAGGACGAUGCGAACUAGCCUCAUCAGCUCAUGUGUCCCUC -81.845

GAGGACGAUGCGAACUAGCCUCAUCAGCUCAUGUGGCCCUC -77.127

GAGGACGAUGCGUACUAGCCUCAUCAGCUCAUGUGACCCUC -79.628

GAGGACGAUGCGUACUAGCCUCAUCAGCUCAUGUGUCCCUC -86.623

GAGGACGAUGCGUACUAGCCUCAUCAGCUCAUGUGGCCCUC -79.604

Table S2.(Continued) The simulation results of 189 mutant RNA aptamer sequences. The mutated positions in the sequences and the ZRNAK scores of three selected aptamers were marked with bold letters.

Type Sequence(5-3) ZRANK Score

Mut

ant s

eque

nce

of S

eq15

GAGGACGAUGCGCACUAGCCUCAUCAGCUCAUGUGACCCUC -72.09

GAGGACGAUGCGCACUAGCCUCAUCAGCUCAUGUGUCCCUC -67.432

GAGGACGAUGCGCACUAGCCUCAUCAGCUCAUGUGGCCCUC -87.531

GAGGACGAUGCGGUCUAGCCUCAUCAGCUCAUGUGCACCUC -67.705

GAGGACGAUGCGGUCUAGCCUCAUCAGCUCAUGUGCUCCUC -59.599

GAGGACGAUGCGGUCUAGCCUCAUCAGCUCAUGUGCGCCUC -66.643

GAGGACGAUGCGGGCUAGCCUCAUCAGCUCAUGUGCACCUC -88.951

GAGGACGAUGCGGGCUAGCCUCAUCAGCUCAUGUGCUCCUC -78.324

GAGGACGAUGCGGGCUAGCCUCAUCAGCUCAUGUGCGCCUC -87.948

GAGGACGAUGCGGCCUAGCCUCAUCAGCUCAUGUGCACCUC -67.698

GAGGACGAUGCGGCCUAGCCUCAUCAGCUCAUGUGCUCCUC -72.326

GAGGACGAUGCGGCCUAGCCUCAUCAGCUCAUGUGCGCCUC -74.909

GAGGACGAUGCGGAAUAGCCUCAUCAGCUCAUGUGCCACUC -81.962

GAGGACGAUGCGGAAUAGCCUCAUCAGCUCAUGUGCCUCUC -78.683

GAGGACGAUGCGGAAUAGCCUCAUCAGCUCAUGUGCCGCUC -74.477

GAGGACGAUGCGGAUUAGCCUCAUCAGCUCAUGUGCCACUC -75.669

GAGGACGAUGCGGAUUAGCCUCAUCAGCUCAUGUGCCUCUC -72.017

GAGGACGAUGCGGAUUAGCCUCAUCAGCUCAUGUGCCGCUC -89.904

GAGGACGAUGCGGAGUAGCCUCAUCAGCUCAUGUGCCACUC -77.132

GAGGACGAUGCGGAGUAGCCUCAUCAGCUCAUGUGCCUCUC -75.5

GAGGACGAUGCGGAGUAGCCUCAUCAGCUCAUGUGCCGCUC -85.028

GAGGACGAUGCGGACAAGCCUCAUCAGCUCAUGUGCCCAUC -78.824

GAGGACGAUGCGGACAAGCCUCAUCAGCUCAUGUGCCCUUC -67.014

GAGGACGAUGCGGACAAGCCUCAUCAGCUCAUGUGCCCGUC -70.113

GAGGACGAUGCGGACGAGCCUCAUCAGCUCAUGUGCCCAUC -77.709

GAGGACGAUGCGGACGAGCCUCAUCAGCUCAUGUGCCCUUC -85.681

GAGGACGAUGCGGACGAGCCUCAUCAGCUCAUGUGCCCGUC -72.912

GAGGACGAUGCGGACCAGCCUCAUCAGCUCAUGUGCCCAUC -79.488

GAGGACGAUGCGGACCAGCCUCAUCAGCUCAUGUGCCCUUC -76.647

GAGGACGAUGCGGACCAGCCUCAUCAGCUCAUGUGCCCGUC -88.449