Single cell whole genome amplification (WGA) with less bias making this kit ideally suited for next generation sequencing applications.

• High sensitivity: Typical yield is greater than 5 µg when starting with a single mammalian cell.
• Reduced amplification bias and no primer artefacts: Primer-free amplification method utilizes a combination of TthPrimPol Primase (to synthesize primers) and Phi29 DNA polymerase.
• Well suited for next generation sequencing: Tested in Illumina and Ion Torrent NGS workflows.
• Easy and reliable
• Insensitive to external DNA contamination

• How does TruePrime™ Technology work?
• Whole genome amplification from 5 cells
• Absence of primer artefacts
• Insensitivity to external DNA contaminations
• Excellent genome coverage

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How does TruePrime technology work?

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Whole genome amplification from 5 cells

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Absence of primer artefacts

1 pg of human genomic DNA (~ 1/6 of the content of one human/mammalian cell) has been amplified using either TruePrime™ (TthPrimPol-based MDA) or random primed MDA reactions. Random primed reactions contain 20% of sequences that cannot be mapped to any organism in sequence databases.

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Insensitivity to external DNA contaminations

Due to the high sensitivity of single cell whole genome amplification techniques there is a considerable danger of contaminating reactions with non-target derived DNA. We therefore recommend to take extreme care when setting up single cell amplification reactions as laid out in the handbook. However, TruePrime™ users have a unique advantage here: TruePrime™ does not accept non-denatured DNA strands as template as readily as random primed MDA reactions. Therefore, contaminations coming in from the environment etc. after the denaturation step will not have a strong influence on the composition of your reaction output. We have shown this behaviour in an experiment where we simulate an external DNA contamination by deliberately adding in yeast DNA to a denatured human DNA template. Yeast DNA is in a thousand-fold excess over the target DNA (1 pg human genomic DNA):

Whereas with TruePrime™ the vast majority of the sequences obtained are target-derived, random primed MDA shows a majority of contaminant-derived sequences.

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Excellent genome coverage

We have amplified and sequenced the yeast genome using TruePrime™ and Illumina technology. Using 2.5 million read pairs we cover 99.4% of the total yeast genome. Shown is the coverage with narrow width of the distribution curve.

A coverage graph of yeast chromosome 7 exemplifies the more even coverage obtained by TruePrime™ MDA (middle blue line represents mean coverage).

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ORDER INFORMATION

Each TruePrime™ Single Cell WGA Kit version 2.0 contains: Buffer L1, DTT, Buffer N, Reaction Buffer, dNTPs, Water, Enzyme 1, Enzyme 2, Control 1, and Control 2. The complete manual is available under the Manual tab. Lucigen is an authorized distributor of Sygnis products in the US. TruePrime™ Single Cell WGA version 2.0 kit is intended for molecular biology use only and in vitro use only. This product is not intended for diagnosis, prevention or treatment of a disease in human beings or animals.

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