全ゲノム領域で均一性が高く偏りのない増幅
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PCR-based WGA can lead to error-prone amplification that results in, for example, single base-pair mutations, STR contractions, and expansions, and also leads to biased and underrepresented loci. In contrast, REPLI-g technology, which uses MDA technology and Phi 29 polymerase, delivers highly uniform amplification across the entire genome with minimal locus bias during amplification — a critical factor for reliable downstream analyses.
Uniform amplification 
Unbiased locus representation
Uniform amplification across the entire genome
The long DNA fragment lengths generated using the highly processive Phi 29 polymerase (see figure Multiple Displacement Amplification (MDA) technology) ensure that REPLI-g amplified DNA covers the whole genome, enabling consistent and unbiased locus representation and minimized mutation rates during amplification (see figures Complete genome coverage and Consistent and accurate whole genome amplification).

Due to unequal amplification of different loci caused by unresolved secondary structures, PCR-based WGA methods such as DOP-PCR or PEP exhibit frequent locus dropout. REPLI-g technology shows highly representative DNA amplification and minimal risk of locus dropout (see figure Highly representative amplification). DNA amplified using REPLI-g provides comparable sequence coverage with unamplified gDNA, making it highly suited for demanding technologies, including next-generation sequencing (see figure Comparable NGS results).

Unbiased locus representation
Phi 29 polymerase is a DNA polymerase with 3'→5' prime exonuclease activity (proofreading activity) that delivers up to 1000-fold higher fidelity compared to Taq DNA polymerase. Supported by the optimized REPLI-g buffer system, Phi 29 polymerase easily solves secondary structures such as hairpin loops, thereby preventing slipping, stoppage, and dissociation of the polymerase during amplification. This enables the generation of DNA fragments up to 100 kb without sequence bias (see figure Unbiased amplification), even from just single cells. Four WGA kits, 2 utilizing MDA technology, including the REPLI-g Single Cell Kit, and 2 PCR-based methods, were tested for sequence representation and locus dropout using the single-cell amplification protocols specific for each kit. Unlike with the REPLI-g Single Cell Kit, single cells analyzed using kits from other suppliers often failed in complete and unbiased sequence representation (see figure Unbiased amplification from a single cell).

注目製品
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REPLI-g Single Cell Kit (24)
シングルセルあるいは限られたサンプル量からの全ゲノム増幅(WGA)
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微量あるいは貴重なサンプルから全ゲノムを均一に増幅
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微量あるいは貴重なサンプルから全ゲノムを均一に増幅
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微量あるいは貴重なサンプルから全ゲノムを均一かつ迅速に増幅
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微量あるいは貴重なサンプルから全ゲノムをハイスループットで増幅(マニュアルあるいは自動化)
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ホルマリン固定パラフィン包埋 (FFPE) 組織切片から直接全ゲノムDNA を増幅
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パンフレット
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Second edition — innovative tools
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キットハンドブック
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For whole genome amplification from single cells, limited samples, or purified genomic DNA
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For fast whole genome amplification from purified genomic DNA, blood, and cells
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For direct whole genome amplification of DNA from formalin-fixed, paraffin-embedded (FFPE) tissue
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For whole genome amplification from purified genomic DNA, blood, and cells
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サイエンティフィック・ポスター
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