Whole Genome Amplification
With the development of whole genome amplification (WGA) techniques, large quantities of DNA can be generated from limited starting quantities. Procedures have been used such as primer extension pre-amplification (PEP) with random 15-mer primers to amplify genomic DNA from single cells or low available amounts of DNA. Other strategies like linker adaptor PCR, tagged PCR, and degenerate oligonucleotide primed PCR (DOP) were designed, improving the yield, but the coverage of the genome in the amplification products was not optimal. Applying multiple displacement amplification (MDA) with random primers and f29 DNA polymerase now ensures accurate whole genome amplification from small amounts of samples.
Whole Genome Amplification (WGA) Kits
Genomic testing and characterization have become an important tool for understanding biological systems. Often, such analysis is hampered by the number of samples to be examined and the availability of sufficient quantities of genomic DNA. This is particularly a problem for rare and archived sources of DNA. The GenomePlex® Whole Genome Amplification (WGA) kits have been developed for use as a high-throughput system for the rapid and highly representative, amplification of genomic DNA from trace amounts of starting material.
The GenomePlex® WGA kits utilize a proprietary amplification technology based upon random fragmentation of genomic DNA and conversion of the resulting small fragments to PCR-amplifiable OmniPlex® Library molecules flanked by universal priming sites. The OmniPlex® library is then amplified using universal oligonucleotide primers and a limited number of cycles. The GenomePlex® WGA kits require nanogram quantities of genomic DNA with yields of 5-10 µg following PCR. This system is suitable with a variety of purified DNA samples including whole blood, blood cards, buccal swabs, plant sources, and formalin-fixed, paraffin-embedded tissue samples.
Single Cell Whole Genome Amplification Strategies
The GenomePlex® Single Cell Whole Genome Amplification Kit (WGA4) is designed to amplify the genome of a single cell resulting in a million-fold amplification yielding microgram quantities of genomic DNA. Traditional single cell whole genome amplification methods yield insufficient quantities with significant biased representation. However, the GenomePlex® technology provides amplification efficiency with WGA DNA Polymerase. The Single Cell WGA kit includes all the reagents necessary for cell lysis and subsequent whole genome amplification.
The Single Cell WGA kit has been validated with numerous downstream applications and is suitable for amplification from a variety of sources including plant, polycarbonate fixed cells, lymphocytes, renal cancer cells, epithelial cells, leukemia cells, and fibroblast amniotic cells. The amplified genome is compatible with many downstream applications including comparative genomic hybridization (CGH), microsatellite/STR analysis, SNP analysis, QPCR, and other genotyping analysis.
SeqPlex-I DNA and RNA Amplification Kits for Illumina® Next Generation Sequencing
The SeqPlex-I DNA and RNA Amplification Kits for whole genome amplification (WGA) and whole transcriptome amplification (WTA) are designed to facilitate Illumina® next-generation sequencing (NGS) from extremely small quantities or degraded/highly fragmented DNA and RNA. Both the WGA and WTA kits utilize a proprietary technology based on random priming of genomic DNA or RNA and conversion of the resulting small fragments to PCR amplifiable library molecules flanked by universal priming sites. The SeqPlex-I WGA and WTA kits allow easy amplification of limited starting samples for convenient direct input onto Illumina® NGS flow cells, enabling you to accelerate your next big discovery.
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