مرکز اطلاعات علمی Scientific Information Database (SID) - Trusted Source for Research and Academic Resources

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Cites:

1

Information Journal Paper

Title

CHALLENGES TO DESIGN AND DEVELOP OF DNA APTAMERS FOR PROTEIN TARGETS. I. OPTIMIZATION OF ASYMMETRIC PCR FOR GENERATION OF A SINGLE STRANDED DNA LIBRARY

Pages

  133-141

Abstract

 APTAMERs, or single stranded oligonucleotides, are produced by systematic evolution of ligands by exponential enrichment, abbreviated as SELEX. In the amplification and regeneration step of SELEX technique, dsDNA is conversed to ssDNA. ASYMMETRIC PCR is one of the methods used for the generation of ssDNA. The purpose of this study was to design a random DNA library for selection of APTAMERs with high affinity for protein targets and develop an efficient ASYMMETRIC PCR amplification. Thus, the influence of factors including annealing temperature, number of amplification cycles, primer ratio, Mg2+ concentration and the presence of a PCR enhancer on the amplification of the desired product were evaluated. Results obtained by agarose gel electrophoresis showed that the annealing temperature of 64 oC, Mg2+ concentration of 0.25 mM, reverse to forward primer ratio of 15: 1, amplification cycle of 25 and the presence L-ectoin as a PCR enhancer with the concentration of 0.4 M were the optimal conditions. Our results supported that the yield of this type of ssDNA production is sufficient for combinatorial screening of APTAMERs.

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    • Cite

    APA: Copy

    TABARZAD, MARYAM, KAZEMI, BAHRAM, VAHIDI, HOSSEIN, ABOOFAZELI, REZA, SHAHHOSSEINI, SORAYA, & NAFISSI VARCHEH, NASTARAN. (2014). CHALLENGES TO DESIGN AND DEVELOP OF DNA APTAMERS FOR PROTEIN TARGETS. I. OPTIMIZATION OF ASYMMETRIC PCR FOR GENERATION OF A SINGLE STRANDED DNA LIBRARY. IRANIAN JOURNAL OF PHARMACEUTICAL RESEARCH (IJPR), 13(SUPPL), 133-141. SID. https://sid.ir/paper/288421/en

    Vancouver: Copy

    TABARZAD MARYAM, KAZEMI BAHRAM, VAHIDI HOSSEIN, ABOOFAZELI REZA, SHAHHOSSEINI SORAYA, NAFISSI VARCHEH NASTARAN. CHALLENGES TO DESIGN AND DEVELOP OF DNA APTAMERS FOR PROTEIN TARGETS. I. OPTIMIZATION OF ASYMMETRIC PCR FOR GENERATION OF A SINGLE STRANDED DNA LIBRARY. IRANIAN JOURNAL OF PHARMACEUTICAL RESEARCH (IJPR)[Internet]. 2014;13(SUPPL):133-141. Available from: https://sid.ir/paper/288421/en

    IEEE: Copy

    MARYAM TABARZAD, BAHRAM KAZEMI, HOSSEIN VAHIDI, REZA ABOOFAZELI, SORAYA SHAHHOSSEINI, and NASTARAN NAFISSI VARCHEH, “CHALLENGES TO DESIGN AND DEVELOP OF DNA APTAMERS FOR PROTEIN TARGETS. I. OPTIMIZATION OF ASYMMETRIC PCR FOR GENERATION OF A SINGLE STRANDED DNA LIBRARY,” IRANIAN JOURNAL OF PHARMACEUTICAL RESEARCH (IJPR), vol. 13, no. SUPPL, pp. 133–141, 2014, [Online]. Available: https://sid.ir/paper/288421/en

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