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Related Concept Videos

DNA Isolation01:24

DNA Isolation

DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...
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RACE - Rapid Amplification of cDNA Ends

Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific primer.
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Related Experiment Video

Updated: May 11, 2026

Primer-Free Aptamer Selection Using A Random DNA Library
11:14

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Published on: July 26, 2010

Single-primer-limited amplification: a method to generate random single-stranded DNA sub-library for aptamer

Chao-Zhu He1, Kun-He Zhang, Ting Wang

  • 1Department of Gastroenterology, First Affiliated Hospital of Nanchang University, Jiangxi Institute of Gastroenterology and Hepatology, Nanchang 330006, China.

Analytical Biochemistry
|May 29, 2013
PubMed
Summary

Single-Primer-Limited Amplification (SPLA) efficiently generates DNA libraries for aptamer selection. This method minimizes by-products, improving DNA amplification for systematic evolution of ligands by exponential enrichment (SELEX).

Keywords:
AptamerMethodologyRandom ssDNA librarySingle-primer-limited amplificationssDNA generation

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Last Updated: May 11, 2026

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Genomics

Background:

  • Polymerase chain reaction (PCR) amplification of single-stranded DNA (ssDNA) libraries is crucial for aptamer selection.
  • Nonspecific hybridization during PCR can lead to by-product amplification, hindering aptamer development.
  • Existing methods struggle to produce clean ssDNA libraries for systematic evolution of ligands by exponential enrichment (SELEX).

Purpose of the Study:

  • To develop a novel method for amplifying random ssDNA libraries free from by-products.
  • To optimize conditions for efficient and specific ssDNA amplification for aptamer selection.

Main Methods:

  • Developed Single-Primer-Limited Amplification (SPLA) using limited primer quantities.
  • Initiated amplification with limited reverse primer for minus-stranded DNA (msDNA).
  • Followed with limited forward primer for plus-stranded DNA (psDNA) and purified via gel excision.

Main Results:

  • SPLA effectively amplified target ssDNA without detectable by-products or nonspecific products.
  • Optimized SPLA produced 16.1 times more psDNA compared to asymmetric PCR.
  • By-product formation correlated with increased template amount and thermal cycles.

Conclusions:

  • SPLA is a simple and feasible method for generating high-quality random ssDNA sub-libraries.
  • This technique significantly improves the efficiency of aptamer selection processes.
  • SPLA offers a robust alternative to conventional PCR for ssDNA library amplification.