By Günter Mayer
This quantity presents protocol references masking contemporary advancements within the aptamer box. in the final decade, aptamers became a growing number of renowned, and their subtle biophysical houses including their skill to be simply transformed and, therefore, tailored to numerous regimens makes them a truly promising classification of compounds. Divided into 3 sections, the e-book covers choice, a sequence of analytical how you can determine biophysical houses of aptamer-target interactions, in addition to numerous purposes of aptamers. Written for the hugely winning Methods in Molecular Biology sequence, chapters contain introductions to their respective issues, lists of the mandatory fabrics and reagents, step by step, conveniently reproducible laboratory protocols, and pointers on troubleshooting and heading off identified pitfalls.
Practical and simple to keep on with, Nucleic Acid Aptamers: choice, Characterization, and Application presents a state of the art precis of modern advancements within the aptamer box and may be a beneficial source for scientists within the existence sciences operating with aptamers as instruments to clarify organic systems.
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Extra resources for Nucleic Acid Aptamers: Selection, Characterization, and Application
Sample text
6. Incubation buffer for treatment: culture medium serum-free. 7. Washing buffer: culture medium serum-free. Developing Aptamers by Cell-Based SELEX 39 8. 6 % sodium dodecyl sulfate (SDS). 9. [γ-32P]ATP (6000 Ci/mmol). 10. Nonspecific competitor: polyinosinic acid (Poly-I). 6 Restriction Fragment Length Polymorphism (RFLP) Analysis 1. 3, 500 mM KCl, 15 mM MgCl2. 2. 02 U/μl Taq polymerase, and DNA template. 3. [γ-32P]ATP (3000 Ci/mmol). 4. 0, 500 mM NaCl, 100 mM MgCl2. 5. Restriction enzymes: RsaI, AluI, HaeIII, HhaI (Invitrogen).
Cut one filter paper and nitrocellulose membrane to the size of the dot blot apparatus. Equilibrate nitrocellulose membrane in aminohexanoic acid buffer for 30 min at room temperature. Afterwards, equilibrate both nitrocellulose membrane and filter paper in SELEX buffer for at least 5 min. 28 Katharina Berg et al. 7. Assemble dot blot apparatus. Therefore, place filter paper beneath the nitrocellulose membrane inside dot blot apparatus. 8. Apply vacuum on dot blot apparatus and wash each well twice with 200 μL SELEX buffer.
Shangguan D, Tang Z, Mallikaratchy P, Xiao Z, Tan W (2007) Optimization and modifications of aptamers selected from live cancer cell lines. Chembiochem 8:603–606 32 Katharina Berg et al. 6. Chu TC, Marks JW 3rd, Lavery LA, Faulkner S, Rosenblum MG, Ellington AD, Levy M (2006) Aptamer:toxin conjugates that specifically target prostate tumor cells. Cancer Res 66:5989–5992 7. Chu TC, Twu KY, Ellington AD, Levy M (2006) Aptamer mediated siRNA delivery. Nucleic Acids Res 34:e73 8. Farokhzad OC, Jon S, Khademhosseini A, Tran TN, Lavan DA, Langer R (2004) Nanoparticle-aptamer bioconjugates: a new approach for targeting prostate cancer cells.