TruePrime™ Single Cell WGA Kit version 2.0 uses a revolutionary novel multiple displacement amplification 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.

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 on dry ice. Upon receipt, the kit should be stored immediately at -20°C in a non-frost-free (constant-temperature) freezer. When stored under these conditions and handled correctly, the products can be kept at least six months after shipping without showing any reduction in performance. For longer periods of time, store the kit at -80°C.

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 4°C for short-term storage or -20°C 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

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.