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TruePrime™ Single Cell WGA Kit

For use with whole cells, where DNA has not yet been extracted from cells

TruePrime™ Single Cell WGA Kit uses a novel and reliable method to achieve accurate genome amplification from single cells or fewer cells. Amplification of a few cells instead of one can improve total coverage breadth if your experimental design allows this.

TruePrime™ Single Cell WGA Kit uses alkaline incubation to allow cell lysis and 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 included – (if DNA already purified select the TruePrime™ WGA kit instead)
  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 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 either a single or a few cells. 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

Supply: TruePrime™ Single Cell WGA Kit is supplied in a single box, with sufficient reagents for either 25 or 100 amplification reactions. The system consists of ten 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™ Single Cell WGA kit handbook.

Shipping and Storage: TruePrime™ Single Cell 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: TruePrime™ Single Cell WGA Kit version 2.0 uses a novel and reliable method to achieve accurate genome amplification from single cells. Dedicated buffers and enzymes deliver microgram quantities of DNA from a single cell or few cells. Up to 50 cells can be amplified with the same protocol. Amplification of a few cells instead of one can improve total coverage breadth if your experimental design allows this. Typical DNA yields from a TruePrime™ Single Cell WGA kit version 2.0 reaction are about 3-4 μg per 50 μl reaction and 3 hours reaction time when starting from a single mammalian cell.
Please note that the apparent yield may change depending on the quantification method chosen. Yields and kinetics will vary if crude or unquantified 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.
TruePrime™ Single Cell WGA Kit version 2.0 uses alkaline incubation to allow cell lysis and 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.

Quality Control: Each batch of TruePrime™ Single Cell 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: TruePrime™ Single Cell WGA kit version 2.0 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. Buffer L1 contains potassium hydroxide: corrosive, harmful. Risk and safety phrases: R22-35. S26-36/37/39-45.

Contents

Description Cap color 25 reactions100 reactions
Reaction BufferYellow1 x 140 μl 1 x 550 μl
Buffer L1Red1 x 70 μl 1 x 250 μl
DTTWhite1 x 50 μl 1 x 50 μl
Buffer NTranslucent 1 x 70 μl 1 x 275 μl
dNTPsGreen 1 x 140 μl1 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
Control 1Amber 1 x 50 μl 1 x 50 μl
Control 2Amber 1 x 50 μl 1 x 50 μl

Note : Product Codes 351025 and 351100 have replaced the previous product codes 350025 and 350100 as we are now manufacturing and distributing this new and improved version of the kit.

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Excellent overall mapping data for TruePrime™-amplified DNA, closest to non-amplified DNA coverage reached.

 

TruePrime WGA reproducbility data

Single Hek293 cells were amplified with different MDA-type single cell amplification methods and coverage compared to the theoretically possible coverage (Poisson) and the coverage in non-amplified DNA from the same Hek293 population. Exactly 12 mio read pairs were mapped to the human genome. Coverage graph for chromosome 3 depicting differences in evenness of coverage between the methods. Note the extreme similarity between nonamplified and TruePrime™.

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