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How does TruePrime™ technology work?

 How does TruePrime™ technology work?


TruePrime™: absence of primer artefacts

 TruePrime™: 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.


TruePrime™ is insensitive 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):

TruePrime™ is insensitive to external DNA contaminations

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


TruePrime™ shows an 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.

TruePrime™ shows an excellent genome coverage

 

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

A coverage graph of yeast chromosome


TruePrime™ Single Cell WGA

Examples of TruePrime™ whole genome amplification

Examples of TruePrime™ whole genome amplification


TruePrime™ WGA

Examples of TruePrime™ whole genome amplification

Examples of TruePrime™ whole genome amplification


TruePrime™ shows an extreme sensitivity

TruePrime™ shows an extreme sensitivity


TruePrime™ produces high yields of DNA from 1 fg of input

TruePrime™ produces high yields of DNA from 1 fg of input


 

TruePrime™ RCA

TruePrime™ RCA Kit: excellent sensitivity from plasmids to BACs

TruePrime™ RCA Kit: excellent sensitivity from plasmids to BACs


 

TruePrime™ RCA Kit: Absence of background amplification

TruePrime™ RCA Kit: Absence of background amplification


 

TruePrime™ RCA Kit: Amplification from bacterial cells

TruePrime™ RCA Kit: Amplification from bacterial cells