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examined and selected. Library capacity for SELEX experiments is practically limited to 10 candidates, whereas, assuming there is a 4-monomeric repertoire from which pools can be created, there are ~1.6 × 10 unique sequences in sequence space limited to a 100-residue matrix, which is clearly beyond experimental capabilities. The library of oligonucleotides must be extremely diverse and not contain linear, incapable of providing a stable spatial arrangement, and double-stranded structures; due to these limitations, oligonucleotide libraries can cover the diversity of only ~10 sequences. This means that existing aptamers may not fully cover the diversity of target molecules or may not have optimal properties due to limitations of the underlying method. To yield the best possible aptamers one must maximize the effectiveness of the discovery process and the library itself.
195:
Ellington and
Szostak demonstrated that chemical synthesis is capable of generating ~10 unique sequences for oligonucleotide libraries in their 1990 paper on in vitro selection, they found that amplification of these synthesized oligonucleotides led to significant loss of pool diversity due to PCR bias and defects in synthesized fragments. The oligonucleotide pool is amplified and a sufficient amount of the initial library is added to the reaction so that there are numerous copies of each individual sequence to minimize the loss of potential binding sequences due to stochastic events. Before the library is introduced to target for incubation and selective retention, the sequence library must be converted to single stranded oligonucleotides to achieve structural conformations with target binding properties.
261:
simplicity, one of the most used methods is using biotinylated reverse primers in the amplification step, after which the complementary strands can be bound to a resin followed by elution of the other strand with lye. Another method is asymmetric PCR, where the amplification step is performed with an excess of forward primer and very little reverse primer, which leads to the production of more of the desired strand. A drawback of this method is that the product should be purified from double stranded DNA (dsDNA) and other left-over material from the PCR reaction. Enzymatic degradation of the unwanted strand can be performed by tagging this strand using a phosphate-probed primer, as it is recognized by enzymes such as
265:. These enzymes then selectively degrade the phosphate tagged strand leaving the complementary strand intact. All of these methods recover approximately 50 to 70% of the DNA. For a detailed comparison refer to the article by Svobodová et al. where these, and other, methods are experimentally compared. In classical SELEX, the process of randomized single stranded library generation, target incubation, and binding sequence elution and amplification described above are repeated until the vast majority of the retained pool consists of target binding sequences, though there are modifications and additions to the procedure that are often used, which are discussed below.
226:
concentrations and homeostatic temperatures are more likely to generate aptamers that can bind in vivo. Another consideration in incubation buffer conditions is non-specific competitors. If there is a high likelihood of non-specific oligonucleotide retention in the reaction conditions, non specific competitors, which are small molecules or polymers other than the SELEX library that have similar physical properties to the library oligonucleotides, can be used to occupy these non-specific binding sites. Varying the relative concentration of target and oligonucleotides can also affect properties of the selected aptamers. If a good
335:) and by machine learning models such as SPOT-RNA, MXfold2 provides the opportunity to assess the ability of sequences in the primary library to fold into complex structures, allowing for the selection of only the most promising sequences from the entire pool. However, these algorithms are low-performance, making them poorly suited for this task. For this reason, algorithms like Ufold from the University of California and AliNA from Nanobiorobots Inc. have been developed, which demonstrate a significant increase in computational speed due to their faster architecture, and can be applied for preliminary
324:, and thus weaken the likelihood of successful binding, particularly when working with small molecules. FRELEX follows a similar overall methodology to SELEX; however, instead of immobilizing the target, the researcher introduces a series of random and blocker oligonucleotides to an immobilization field before introduction to the target. This allows the researcher to better target small molecules that may be lost during partitioning. It also can be used in some circumstances to select an aptamer library without knowing the target.
274:
negative selection can be used where the library is incubated with target immobilization matrix components and unbound sequences are retained. Negative selection can also be used to eliminate sequences that bind target-like molecules or cells by incubating the oligonucleotide library with small molecule target analogs, undesired cell types, or non-target proteins and retaining the unbound sequences.
33:
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through elution concentration estimations via 260 nm wavelength absorbance or fluorescent labeling of oligonucleotides. As the SELEX reaction approaches completion, the fraction of the oligonucleotide library that binds target approaches 100%, such that the number of eluted molecules approaches the total oligonucleotide input estimate, but may converge at a lower number.
41:
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oligonucleotide-target binding. There are several considerations for this target incubation step, including the target immobilization method and strategies for subsequent unbound oligonucleotide separation, incubation time and temperature, incubation buffer conditions, and target versus oligonucleotide concentrations. Examples of target immobilization methods include
244:
sequences are then eluted by creating denaturing conditions that promote oligonucleotide unfolding or loss of binding conformation including flowing in deionized water, using denaturing solutions containing urea and EDTA, or by applying high heat and physical force. Upon elution of bound sequences, the retained oligonucleotides are
391:. Additionally, SELEX has been utilized to obtain highly specific catalytic DNA or DNAzymes. Several metal-specific DNAzymes have been reported including the GR-5 DNAzyme (lead-specific), the CA1-3 DNAzymes (copper-specific), the 39E DNAzyme (uranyl-specific) and the NaA43 DNAzyme (sodium-specific).
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Recently, SELEX has expanded to include the use of chemically modified nucleotides. These chemically modified oligonucleotides offer many potential advantages for selected aptamers including greater stability and nuclease resistance, enhanced binding for select targets, expanded physical properties -
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Once the oligonucleotide library has been incubated with target for sufficient time, unbound oligonucleotides are washed away from immobilized target, often using the incubation buffer so that specifically bound oligonucleotides are retained. With unbound sequences washed away, the specifically bound
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SELEX has been used to develop a number of aptamers that bind targets interesting for both clinical and research purposes. Nucleotides with chemically modified sugars and bases have been incorporated into SELEX reactions to increase the chemical diversity at each base, expanding the possibilities for
185:
Aptamers have emerged as a novel category in the field of bioreceptors due to their wide applications ranging from biosensing to therapeutics. Several variations of their screening process, called SELEX have been reported which can yield sequences with desired properties needed for their final use.
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Most modern aptamer selection methods strive to improve the conventional SELEX aptamer search method. Despite the publication of various methods aimed at increasing the affinity and specificity of aptamers, experimental approaches face limitations in the number and variety of sequences that can be
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to allow nucleotides to approach the target and increase the chance of a specific binding event. Alternatively, if the desired aptamer function is in vivo protein or whole cell binding for potential therapeutic or diagnostic application, incubation buffer conditions similar to in vivo plasma salt
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Immediately prior to target introduction, the single stranded oligonucleotide library is often heated and cooled slowly to renature oligonucleotides into thermodynamically stable secondary and tertiary structures. Once prepared, the randomized library is incubated with immobilized target to allow
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In order to increase the specificity of aptamers selected by a given SELEX procedure, a negative selection, or counter selection, step can be added prior to or immediately following target incubation. To eliminate sequences with affinity for target immobilization matrix components from the pool,
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One of the most critical steps in the SELEX procedure is obtaining single stranded DNA (ssDNA) after the PCR amplification step. This will serve as input for the next cycle so it is of vital importance that all the DNA is single stranded and as little as possible is lost. Because of the relative
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To track the progress of a SELEX reaction, the number of target bound molecules, which is equivalent to the number of oligonucleotides eluted, can be compared to the estimated total input of oligonucleotides following elution at each round. The number of eluted oligonucleotides can be estimated
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Some SELEX reactions can generate probes that are dependent on primer binding regions for secondary structure formation. There are aptamer applications for which a short sequence, and thus primer truncation, is desirable. An advancement on the original method allows an RNA library to omit the
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The first step of SELEX involves the synthesis of fully or partially randomized oligonucleotide sequences of some length flanked by defined regions which allow PCR amplification of those randomized regions and, in the case of RNA SELEX, in vitro transcription of the randomized sequence. While
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220:
and incubated with the oligonucleotide library on culture plates. Incubation buffer conditions are altered based on the intended target and desired function of the selected aptamer. For example, in the case of negatively charged small molecules and proteins, high salt buffers are used for
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for the selected aptamer is not a concern, then an excess of target can be used to increase the probability that at least some sequences will bind during incubation and be retained. However, this provides no selective pressure for high
158:
to prepare for subsequent rounds of selection in which the stringency of the elution conditions can be increased to identify the tightest-binding sequences. A caution to consider in this method is that the selection of extremely high,
371:(VEGF). Moreover, SELEX has been used to select high-affinity aptamers for complex targets such as tumor cells, tumor exosomes, or tumor tissue. Clinical uses of the technique are suggested by aptamers that bind
308:
The genetic alphabet, and thus possible aptamers, is also expanded using unnatural base pairs the use of these unnatural base pairs was applied to SELEX and high affinity DNA aptamers were generated.
1920:
Nagano M, Toda T, Makino K, Miki H, Sugizaki Y, Tomizawa H, et al. (December 2022). "Discovery of a Highly
Specific Anti-methotrexate (MTX) DNA Aptamer for Antibody-Independent MTX Detection".
320:
to allow the selection of aptamers without immobilizing the target or the oligonucleotide library. Immobilization is a necessary component of SELEX; however, it has the potential to inhibit key
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Mercey R, Lantier I, Maurel MC, Grosclaude J, Lantier F, Marc D (November 2006). "Fast, reversible interaction of prion protein with RNA aptamers containing specific sequence patterns".
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to DNA in the case of RNA or modified base selections, or simply collected for amplification in the case of DNA SELEX. These DNA templates from eluted sequences are then amplified via
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Ulrich H, Trujillo CA, Nery AA, Alves JM, Majumder P, Resende RR, Martins AH (September 2006). "DNA and RNA aptamers: from tools for basic research towards therapeutic applications".
2964:
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Lubin AA, Hunt BV, White RJ, Plaxco KW (March 2009). "Effects of probe length, probe geometry, and redox-tag placement on the performance of the electrochemical E-DNA sensor".
36:
A schematic of the major phases in a SELEX experiment. This cycle, may be repeated up to 20 times over a period lasting weeks, though some methods require far fewer cycles.
619:
Blackwell TK, Weintraub H (November 1990). "Differences and similarities in DNA-binding preferences of MyoD and E2A protein complexes revealed by binding site selection".
2826:
Ferreira CS, Matthews CS, Missailidis S (2006). "DNA aptamers that bind to MUC1 tumour marker: design and characterization of MUC1-binding single-stranded DNA aptamers".
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binding affinity entities may not in fact improve specificity for the target molecule. Off-target binding to related molecules could have significant clinical effects.
1686:, Penner, Gregory & CA, "United States Patent: 10415034 - Method for the selection of aptamers for unbound targets", issued September 17, 2019
122:
The process begins with the synthesis of a very large oligonucleotide library, consisting of randomly generated sequences of fixed length flanked by constant
2929:
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1480:
Pinheiro VB, Taylor AI, Cozens C, Abramov M, Renders M, Zhang S, Chaput JC, Wengel J, Peak-Chew SY, McLaughlin SH, Herdewijn P, Holliger P (April 2012).
235:, which requires the oligonucleotide library to be in excess so that there is competition between unique sequences for available specific binding sites.
3172:
Umrao S, Jain V, Chakraborty B, Roy R (August 2018). "Protein-induced fluorescence enhancement as aptamer sensing mechanism for thrombin detection".
2620:
Mayer G, Ahmed MS, Dolf A, Endl E, Knolle PA, Famulok M (December 2010). "Fluorescence-activated cell sorting for aptamer SELEX with cell mixtures".
1635:
Kimoto M, Yamashige R, Matsunaga K, Yokoyama S, Hirao I (May 2013). "Generation of high-affinity DNA aptamers using an expanded genetic alphabet".
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and converted to single stranded DNA, RNA, or modified base oligonucleotides, which are used as the initial input for the next round of selection.
2433:"RNA aptamers to the adenosine moiety of S-adenosyl methionine: structural inferences from variations on a theme and the reproducibility of SELEX"
2347:
Nasaev SS, Mukanov AR, Kuznetsov II, Veselovsky AV (December 2023). "AliNA - a deep learning program for RNA secondary structure prediction".
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positions with four possibilities (A,T,C,G) at each position). The sequences in the library are exposed to the target ligand - which may be a
1163:"Systematic evolution of a DNA aptamer binding to rat brain tumor microvessels. selective targeting of endothelial regulatory protein pigpen"
1877:
Tuerk C, Gold L (August 1990). "Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase".
492:
Tuerk C, Gold L (August 1990). "Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase".
1109:"Short bioactive Spiegelmers to migraine-associated calcitonin gene-related peptide rapidly identified by a novel approach: tailored-SELEX"
134:, while a small number of random regions are expected to bind specifically to the chosen target. For a randomly generated region of length
1253:
Stoltenburg R, Reinemann C, Strehlitz B (October 2007). "SELEX--a (r)evolutionary method to generate high-affinity nucleic acid ligands".
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These developed aptamers have seen diverse application in therapies for macular degeneration and various research applications including
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115:(cyclic amplification and selection of targets) SELEX was first introduced in 1990. In 2015, a special issue was published in the
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Iwagawa T, Ohuchi SP, Watanabe S, Nakamura Y (January 2012). "Selection of RNA aptamers against mouse embryonic stem cells".
116:
1963:
Gotrik MR, Feagin TA, Csordas AT, Nakamoto MA, Soh HT (September 2016). "Advancements in
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17:
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107:. Although SELEX has emerged as the most commonly used name for the procedure, some researchers have referred to it as
2000:"Multiparameter Particle Display (MPPD): A Quantitative Screening Method for the Discovery of Highly Specific Aptamers"
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48:. The aptamer surface and backbone are shown in yellow. Biotin (spheres) fits snugly into a cavity of the RNA surface.
2171:"RNA secondary structure prediction using an ensemble of two-dimensional deep neural networks and transfer learning"
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131:
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Sefah K, Shangguan D, Xiong X, O'Donoghue MB, Tan W (June 2010). "Development of DNA aptamers using Cell-SELEX".
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155:
2722:"ADAPT identifies an ESCRT complex composition that discriminates VCaP from LNCaP prostate cancer cell exosomes"
2563:"A tenascin-C aptamer identified by tumor cell SELEX: systematic evolution of ligands by exponential enrichment"
216:
Recently, SELEX reactions have been developed where the target is whole cells, which are expanded near complete
222:
2771:"Poly-ligand profiling differentiates trastuzumab-treated breast cancer patients according to their outcomes"
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constant primer regions, which can be difficult to remove after the selection process because they stabilize
205:
151:
72:
1290:"In vitro selection of a 7-methyl-guanosine binding RNA that inhibits translation of capped mRNA molecules"
568:
Ellington AD, Szostak JW (August 1990). "In vitro selection of RNA molecules that bind specific ligands".
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1539:"An unnatural base pair system for efficient PCR amplification and functionalization of DNA molecules"
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Franciscis V (ed.).
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672:"Cyclic amplification and selection of targets (CASTing) for the myogenin consensus binding site"
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3115:"In vitro selection of a sodium-specific DNAzyme and its application in intracellular sensing"
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RNA and DNA secondary structure prediction by dynamic programming algorithms such as RNAfold (
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2665:"Plasma Exosome Profiling of Cancer Patients by a Next Generation Systems Biology Approach"
848:"Aptamers selected for higher-affinity binding are not more specific for the target ligand"
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or target capture on paramagnetic beads. The bound sequences are eluted and amplified by
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1820:"Aptamers as biomarkers for neurological disorders. Proof of concept in transgenic mice"
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1588:"Highly specific unnatural base pair systems as a third base pair for PCR amplification"
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428: – Method for identifying the sequence-specific target site of a DNA-binding domain
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2228:"RNA secondary structure prediction using deep learning with thermodynamic integration"
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138:, the number of possible sequences in the library using conventional DNA or RNA is 4 (
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398:, fluorescent labeling of proteins and cells, and selective enzyme inhibition.
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1433:"In vitro selection using a dual RNA library that allows primerless selection"
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of extremely high binding affinity to a variety of target ligands, including
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like increased hydrophobicity, and more diverse structural conformations.
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Technique for producing oligonucleotides that specifically bind to a target
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416: – DNA oligonucleotides that can perform a specific chemical reaction
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1448:
1124:
459:
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2978:
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410: – Oligonucleotide or peptide molecules that bind specific targets
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104:
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364:
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150:- and those that do not bind the target are removed, usually by
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100:
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84:
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1107:
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specific and sensitive binding, or increasing stability in
119:
in the honor of quarter century of the discovery of SELEX.
2390:
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that are unstable when formed by the random region alone.
2283:
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53:
Systematic evolution of ligands by exponential enrichment
18:
Systematic Evolution of Ligands by Exponential Enrichment
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SELEX variants and alternative aptamer selection methods
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Combinatorial Chemistry & High Throughput Screening
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387:has been approved by the FDA for treatment of
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721:"SELEX: How It Happened and Where It will Go"
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485:
483:
481:
479:
477:
475:
473:
471:
469:
670:Wright WE, Binder M, Funk W (August 1991).
3015:"In vitro selection of self-cleaving DNAs"
1702:Kohlberger M, Gadermaier G (August 2021).
1678:
1676:
1674:
975:"Microtitre Plate Based Cell-SELEX Method"
239:Binding sequence elution and amplification
111:(selected and amplified binding site) and
3408:
3398:
3380:
3345:
3291:
3242:
3232:
3148:
3138:
3089:
3079:
3030:
2924:
2922:
2902:
2871:"RNA mimics of green fluorescent protein"
2869:Paige JS, Wu KY, Jaffrey SR (July 2011).
2802:
2745:
2696:
2596:
2586:
2521:
2519:
2456:
2407:
2323:
2313:
2303:
2259:
2202:
2121:
2064:
2023:
1853:
1843:
1794:
1784:
1727:
1611:
1562:
1513:
1456:
1323:
1313:
1178:
1132:
998:
895:
893:
891:
871:
822:
812:
752:
695:
383:, and a VEGF-binding aptamer trade-named
44:Structure of an RNA aptamer specific for
3266:Mondragón E, Maher LJ (September 2015).
852:Journal of the American Chemical Society
448:Zeitschrift für Urologie und Nephrologie
39:
438:
2956:
2931:Ophthalmology Clinics of North America
2049:Current Protocols in Molecular Biology
1708:Biotechnology and Applied Biochemistry
1212:Analytical and Bioanalytical Chemistry
347:The technique has been used to evolve
7:
3326:Computational Biology and Chemistry
1288:Haller AA, Sarnow P (August 1997).
1167:The Journal of Biological Chemistry
940:Current Opinion in Chemical Biology
91:that specifically bind to a target
3338:10.1016/j.compbiolchem.2006.10.002
3322:"A mathematical analysis of SELEX"
369:vascular endothelial growth factor
95:or ligands. These single-stranded
25:
3174:Sensors and Actuators B: Chemical
973:Shorie M, Kaur H (October 2018).
130:ends. The constant ends serve as
2096:Kosuri S, Church GM (May 2014).
316:FRELEX was developed in 2016 by
300:Chemically modified nucleotides
725:Journal of Molecular Evolution
676:Molecular and Cellular Biology
278:Tracking selection progression
117:Journal of Molecular Evolution
1:
3032:10.1016/s1074-5521(96)90170-2
2431:Burke DH, Gold L (May 1997).
2051:. Chapter 24 (1): Unit 24.2.
1965:Accounts of Chemical Research
339:analysis of these libraries.
318:NeoVentures Biotechnology Inc
269:Negative or counter selection
3221:Journal of Nanobiotechnology
2992:10.1016/1074-5521(94)90014-0
2057:10.1002/0471142727.mb2402s88
1977:10.1021/acs.accounts.6b00283
1934:10.1021/acs.analchem.2c04182
1845:10.1371/journal.pone.0190212
1786:10.1371/journal.pone.0205460
1267:10.1016/j.bioeng.2007.06.001
1083:10.1016/j.biochi.2011.10.017
814:10.1371/journal.pone.0134403
103:are commonly referred to as
914:10.1016/j.ymeth.2016.04.020
426:Bacterial one-hybrid system
3475:
2795:10.1038/s41467-018-03631-z
2540:10.2174/138620706778249695
2298:(3). Bioinformatics: e14.
2252:10.1038/s41467-021-21194-4
2195:10.1038/s41467-019-13395-9
952:10.1016/j.cbpa.2008.06.028
287:Caveats and considerations
83:of either single-stranded
3194:10.1016/j.snb.2018.04.039
2943:10.1016/j.ohc.2006.05.008
2494:10.1007/s00705-006-0790-3
2305:10.1101/2020.08.17.254896
1224:10.1007/s00216-012-6183-4
745:10.1007/s00239-015-9705-9
2146:"ViennaRNA Web Services"
1255:Biomolecular Engineering
719:Gold L (December 2015).
446:Hak-Hagir A (1978). "".
208:columns, nitrocellulose
3400:10.1073/pnas.1605086113
3140:10.1073/pnas.1420361112
3081:10.1073/pnas.0607875104
3019:Chemistry & Biology
2980:Chemistry & Biology
2945:(inactive 2024-09-12).
2895:10.1126/science.1207339
2588:10.1073/pnas.2136683100
1899:10.1126/science.2200121
1506:10.1126/science.1217622
1315:10.1073/pnas.94.16.8521
991:10.21769/BioProtoc.3051
641:10.1126/science.2174572
514:10.1126/science.2200121
206:affinity chromatography
152:affinity chromatography
73:combinatorial chemistry
59:), also referred to as
3272:Nucleic Acids Research
2726:Nucleic Acids Research
2634:10.1038/nprot.2010.163
2449:10.1093/nar/25.10.2020
2437:Nucleic Acids Research
2361:10.1002/minf.202300113
2292:Nucleic Acids Research
2016:10.1002/anie.201608880
1592:Nucleic Acids Research
1543:Nucleic Acids Research
1437:Nucleic Acids Research
1180:10.1074/jbc.M100347200
1113:Nucleic Acids Research
420:Anti-thrombin aptamers
148:small organic compound
49:
37:
3234:10.1186/1477-3155-8-8
2775:Nature Communications
2349:Molecular Informatics
2232:Nature Communications
2175:Nature Communications
1364:10.1038/nprot.2010.66
688:10.1128/mcb.11.8.4104
363:and proteins such as
210:binding assay filters
43:
35:
2482:Archives of Virology
2315:10.1093/nar/gkab1074
2150:rna.tbi.univie.ac.at
1922:Analytical Chemistry
1637:Nature Biotechnology
1398:Analytical Chemistry
389:macular degeneration
294:secondary structures
3454:Genetics techniques
3391:2016PNAS..11312076S
3375:(43): 12076–12081.
3186:2018SeAcB.267..294U
3131:2015PNAS..112.5903T
3072:2007PNAS..104.2056L
2887:2011Sci...333..642P
2787:2018NatCo...9.1219D
2738:10.1093/nar/gkaa034
2681:2017NatSR...742741D
2579:2003PNAS..10015416D
2244:2021NatCo..12..941S
2187:2019NatCo..10.5407S
1928:(49): 17255–17262.
1891:1990Sci...249..505T
1836:2018PLoSO..1390212L
1777:2018PLoSO..1305460K
1604:10.1093/nar/gkr1068
1498:2012Sci...336..341P
1306:1997PNAS...94.8521H
1039:10.1021/bi00002a033
805:2015PLoSO..1034403S
737:2015JMolE..81..140G
633:1990Sci...250.1104B
582:1990Natur.346..818E
506:1990Sci...249..505T
246:reverse-transcribed
214:paramagnetic beads.
3284:10.1093/nar/gkv702
2669:Scientific Reports
2355:(12): e202300113.
2114:10.1038/nmeth.2918
1555:10.1093/nar/gkn956
1449:10.1093/nar/gkl463
1125:10.1093/nar/gng130
263:Lambda exonuclease
68:in vitro evolution
62:in vitro selection
50:
38:
3459:Molecular biology
2840:10.1159/000096085
2689:10.1038/srep42741
2628:(12): 1993–2004.
2004:Angewandte Chemie
1885:(4968): 505–510.
1410:10.1021/ac802317k
864:10.1021/ja060952q
627:(4984): 1104–10.
199:Target incubation
77:molecular biology
16:(Redirected from
3466:
3422:
3412:
3402:
3384:
3359:
3349:
3306:
3305:
3295:
3263:
3257:
3256:
3246:
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3206:
3205:
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3110:
3104:
3103:
3093:
3083:
3051:
3045:
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3010:
3004:
3003:
2975:
2969:
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2954:
2926:
2917:
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2906:
2866:
2860:
2859:
2823:
2817:
2816:
2806:
2766:
2760:
2759:
2749:
2732:(8): 4013–4027.
2717:
2711:
2710:
2700:
2660:
2654:
2653:
2622:Nature Protocols
2617:
2611:
2610:
2600:
2590:
2573:(26): 15416–21.
2558:
2552:
2551:
2523:
2514:
2513:
2488:(11): 2197–214.
2477:
2471:
2470:
2460:
2428:
2422:
2421:
2411:
2387:
2381:
2380:
2344:
2338:
2337:
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2307:
2289:
2280:
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2217:
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2206:
2166:
2160:
2159:
2157:
2156:
2142:
2136:
2135:
2125:
2093:
2087:
2086:
2068:
2044:
2038:
2037:
2027:
1995:
1989:
1988:
1971:(9): 1903–1910.
1960:
1954:
1953:
1917:
1911:
1910:
1874:
1868:
1867:
1857:
1847:
1815:
1809:
1808:
1798:
1788:
1771:(10): e0205460.
1756:
1750:
1749:
1731:
1720:10.1002/bab.2244
1714:(5): 1771–1792.
1699:
1693:
1692:
1691:
1687:
1680:
1669:
1668:
1649:10.1038/nbt.2556
1632:
1626:
1625:
1615:
1583:
1577:
1576:
1566:
1534:
1528:
1527:
1517:
1477:
1471:
1470:
1460:
1428:
1422:
1421:
1393:
1384:
1383:
1352:Nature Protocols
1347:
1338:
1337:
1327:
1317:
1285:
1279:
1278:
1250:
1244:
1243:
1207:
1201:
1200:
1182:
1158:
1147:
1146:
1136:
1119:(21): 130e–130.
1104:
1095:
1094:
1066:
1051:
1050:
1022:
1013:
1012:
1002:
970:
964:
963:
935:
926:
925:
897:
886:
885:
875:
843:
837:
836:
826:
816:
784:
767:
766:
756:
731:(5–6): 140–143.
716:
710:
709:
699:
667:
661:
660:
616:
610:
609:
590:10.1038/346818a0
576:(6287): 818–22.
565:
526:
525:
500:(4968): 505–10.
489:
464:
463:
443:
233:binding affinity
228:binding affinity
223:charge screening
81:oligonucleotides
21:
3474:
3473:
3469:
3468:
3467:
3465:
3464:
3463:
3439:
3438:
3430:
3425:
3362:
3319:
3315:
3313:Further reading
3310:
3309:
3278:(15): 7544–55.
3265:
3264:
3260:
3214:
3213:
3209:
3171:
3170:
3166:
3112:
3111:
3107:
3053:
3052:
3048:
3025:(12): 1039–46.
3012:
3011:
3007:
2977:
2976:
2972:
2955:
2928:
2927:
2920:
2881:(6042): 642–6.
2868:
2867:
2863:
2825:
2824:
2820:
2768:
2767:
2763:
2719:
2718:
2714:
2662:
2661:
2657:
2619:
2618:
2614:
2560:
2559:
2555:
2525:
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2517:
2479:
2478:
2474:
2430:
2429:
2425:
2389:
2388:
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2341:
2287:
2282:
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2277:
2225:
2224:
2220:
2168:
2167:
2163:
2154:
2152:
2144:
2143:
2139:
2095:
2094:
2090:
2046:
2045:
2041:
1997:
1996:
1992:
1962:
1961:
1957:
1919:
1918:
1914:
1876:
1875:
1871:
1830:(1): e0190212.
1817:
1816:
1812:
1758:
1757:
1753:
1701:
1700:
1696:
1689:
1682:
1681:
1672:
1634:
1633:
1629:
1598:(6): 2793–806.
1585:
1584:
1580:
1536:
1535:
1531:
1492:(6079): 341–4.
1479:
1478:
1474:
1430:
1429:
1425:
1395:
1394:
1387:
1349:
1348:
1341:
1287:
1286:
1282:
1252:
1251:
1247:
1209:
1208:
1204:
1173:(19): 16464–8.
1160:
1159:
1150:
1106:
1105:
1098:
1068:
1067:
1054:
1024:
1023:
1016:
972:
971:
967:
937:
936:
929:
899:
898:
889:
858:(24): 7929–37.
845:
844:
840:
799:(7): e0134403.
786:
785:
770:
718:
717:
713:
669:
668:
664:
618:
617:
613:
567:
566:
529:
491:
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467:
445:
444:
440:
435:
404:
353:small molecules
345:
314:
302:
289:
280:
271:
258:
256:Obtaining ssDNA
241:
201:
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183:
28:
23:
22:
15:
12:
11:
5:
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3428:External links
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3125:(19): 5903–8.
3105:
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2861:
2834:(6): 289–301.
2828:Tumour Biology
2818:
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2515:
2472:
2443:(10): 2020–4.
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2108:(5): 499–507.
2102:Nature Methods
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2066:2027.42/143624
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2010:(3): 744–747.
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