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Target Cell Pre-enrichment and Whole Genome Amplification for Single Cell Downstream Characterization
Published on: May 15, 2018
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Deterministic Whole-Genome Amplification of Single Cells
Zbigniew Tadeusz Czyż1, Christoph A Klein2,3
1Project Group, Personalized Tumor Therapy, Fraunhofer Institute for Toxicology and Experimental Medicine (ITEM), Regensburg, Germany.
Methods in Molecular Biology (Clifton, N.J.)
|September 17, 2015
Summary
This chapter details a single-cell whole genome amplification (WGA) method, Ampli1™ WGA Kit, for uniform DNA amplification from single cells. This PCR-based technique enables high-quality genomic material for various downstream genetic assays.
Area of Science:
- Molecular Biology
- Genomics
- Biotechnology
Background:
- Single-cell analysis requires robust methods for whole genome amplification (WGA).
- Existing WGA techniques may suffer from non-uniform amplification or low efficiency.
- The Ampli1™ WGA Kit provides a commercial solution based on a previously published method.
Purpose of the Study:
- To describe a ligation-mediated adaptor linker PCR-based whole genome amplification protocol for single cells.
- To highlight the advantages of nonrandom genome fragmentation and high priming efficiency.
- To provide high-quality amplified DNA suitable for diverse downstream genomic assays.
Main Methods:
- Utilizes a ligation-mediated adaptor linker PCR approach for whole genome amplification (WGA).
- Employs a nonrandom primer design and genome fragmentation mechanism.
- Optimized for amplifying DNA from low-input quantities, specifically single cells.
Main Results:
- Achieves comprehensive and uniform amplification of DNA from single cells.
- Generates high-quality amplified genomic material in sufficient quantities.
- Demonstrates suitability for various downstream applications including Sanger sequencing, RFLP, qPCR, and array CGH.
Conclusions:
- The described WGA method offers a reliable approach for single-cell genome analysis.
- The nonrandom fragmentation and high priming efficiency contribute to superior amplification quality.
- This technique is valuable for diagnostic assays and a wide range of genomic studies.
