DNA amplification reagent kit Ampli1 WGA PLUS

DNA polymeraseTaq DNA polymeraseNGS

The Ampli1 WGA PLUS Kit has been developed and optimized for balanced and complete amplification of the total DNA content of a single human cell. Input DNA (from one single cell and up to 1000 cells in ~1 μI of PBS) may be derived from fresh/live and/or fixed cells (PFA2%, CellSave, CellRescue tubes). The kit can also be used to amplify up to 10 ng of genomic DNA (e.g. from tissue or FFPE samples). Following cell lysis, DNA is digested with a restriction enzyme and adaptors are ligated onto the DNA fragments. This ensures that, for any given locus in the genome, the corresponding alleles are amplified as genomic fragments with same-length and almost identical GC content, thus drastically reducing allelic drop-out due to different amplification efficiency of the original DNA molecules. Amplification, performed in the same tube, is mediated by a single highly specific universal PCR primer for all the fragments and uses a DNA polymerase with 3’→5’ exonuclease proofreading activity, with 6.5X greater fidelity over standard Taq DNA polymerase. The output of the Ampli1 WGA protocol is a well-balanced library of fragments of about 0.2‐2 kb representing the entire genome. Many different types of downstream analysis procedures are compatible with Ampli1 WGA PLUS product, such as variant detection by Sanger or Next Generation sequencing (NGS), microsatellite or other PCR‐based genotyping analysis, metaphase CGH, array CGH and genome‐wide copy number analysis by NGS. Two dedicated downstream kits, Ampli1 OncoSeek Panel and Ampli1 LowPass Kit for Illumina,