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TruePrime™ WGA kit

Whole Genome Amplification Kit for Purified DNA

For use with DNA that has already been pre-extracted from cells

TruePrime™ consists of a set of reagents for multiple displacement amplification of genomic DNA from purified material, with a minimal level of bias. TruePrime™ WGA Kit uses alkaline incubation to allow DNA denaturation of genomic DNA with very low DNA fragmentation. This results in amplified DNA with high integrity and fragment length, so that most of the sequences are uniformly represented.

Features and Benefits:

  1. Cell lysis buffer not included – (select TruePrime™ single cell WGA kit if DNA is not purified)
  2. Primer-free method – TthPrimPol synthesizes the primers for Phi29 DNA polymerase
  3. No primer artefacts – No random extension of primer dimers
  4. Insensitive to external DNA contamination
  5. Works with all types of genomes
  6. Reduced amplification bias in genome coverage – reliable results
  7. Ideally suited for Next Generation Sequencing

Trueprime Whole Genome Amplification process diagram

TruePrime™ technology further information

TruePrime™ technology uses a revolutionary novel multiple displacement amplification (MDA) method based on the combination of the recently discovered DNA primase ‘TthPrimPol’ and the extremely processive and high-fidelity Phi29 DNA polymerase to amplify uniformly total genomic DNA from purified material. The extraordinary strand displacement capacity of the Phi29 DNA polymerase allows TthPrimPol to generate new primers on the displaced strands that are extended by Phi29 DNA pol, resulting in exponential isothermal DNA amplification.

TruePrime multiple displacement amplification in 6 steps

Specifications

Supply: The TruePrime™ WGA kit is supplied in a single box, with sufficient reagents for either 25 or 100 amplification reactions. The system consists of seven components, including the enzymes and buffers to perform the amplification reactions. Contents are provided in clear, screw-cap tubes with colour-coded lids as referenced in the TruePrime™ WGA kit handbook.

Shipping and Storage: The TruePrime™ WGA kit is shipped in dry ice. Upon receipt, the kit should be stored immediately at -20°C in a non-frost-free (constant temperature) freezer. If stored correctly the product can be kept for at least 24 months after shipping without displaying any reduction in performance. For longer storage periods we recommend a temperature of -80ºC.

Technical specifications: The TruePrime™ WGA kit uses a novel and reliable method to achieve accurate genome amplification from purified material. Dedicated buffers and enzymes deliver microgram quantities of DNA. Typical DNA yields from a TruePrime™ WGA kit reaction are above 5 μg per 50 μl reaction when starting from 1 ng of genomic DNA. Yields and kinetics will vary if crude or un-quantified samples are amplified. Reactions without input DNA (no template controls) do not produce any amplification product during 3 hour reaction times. Mean product length is greater than 10 kb. Store amplified DNA at 4ºC for short-term storage or -20ºC for long-term storage.
The TruePrime™ WGA kit uses alkaline incubation to allow DNA denaturation of genomic DNA with very low DNA fragmentation, resulting in amplified DNA with high integrity and maximizing the length of the amplification fragments, so that most of the sequences are uniformly represented.

Quality Control: Each batch of TruePrime™ WGA kit is tested against predetermined specifications to ensure consistent product quality. Enzymes used in the kit have been tested separately to ensure adherence to specifications.

Intended use: The TruePrime™ WGA 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 humans beings or animals.

Patent: Patent protection for TthPrimPol and methods associated with its use is pending.

Safety information: Safety data sheets are available for download.
This material may contain substances or activities that are harmful to human health. It should not be ingested, inhaled, or brought into contact with skin, and handled with appropriate care in accordance with the principles of good laboratory practices.

Kit contents


Description
Cap color25 reactions 100 reactions
Reaction Buffer Yellow 1 x 140 μl1 x 550 μl
Buffer DRed1 x 70 μl 1 x 275 μl
Buffer NTranslucent1 x 70 μl 1 x 275 μl
dNTPsGreen 1 x 140 μl 1 x 550 μl
H2OBlue 1 x 1500 μl 2 x 1500 μl
Enzyme 1Purple 1 x 140 μl 1 x 550 μl
Enzyme 2Orange 1 x 20 μl 1 x 80 μl
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DNA from 4 single human HEK293 cells was amplified following the TruePrimeTM protocol for 3 or 6 h, subjected to library prep and sequencing (Illumina HiSeq), and 5 million read pairs were mapped onto the human genome.

TruePrime WGA reproducbility data

Left: Shown are comparisons of chromosome 3 and 4 coverage to non-amplified HEK293 cell DNA from the same cells. Note the evenness of coverage, and the high similarity between single cells. Right: Circos plot showing the whole genome coverage of the 4 replicates (blue=non-amplified; green=amplified).

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