The Ampli1™ Whole Genome Amplification (WGA) Kit has been developed and optimized for balanced and complete amplification of the total DNA content of a single cell. The output of the Ampli1 WGA protocol is a well-balanced library representing the entire genome. This DNA library is ideally suited for downstream analysis of both SNPs and CNAs in single cells, due to its low rate of preferential amplification.
Ampli1 WGA kit is the only fully “non random” whole genome amplification kit for single, human cell. It provides a robust and reproducible amplification efficiency on every locus and on each one of the two alleles reducing preferential amplification and achieving the lower ADO rate compared to other methods (Binder V et al, Hum Mutat. 2014 - Huang L et al, Annu Rev Genomics Hum Genet. 2015 - Babayan A et al, Oncotarget. 2016 - Borgström E et al, PLoS One. 2017 - Normand E et al, Prenat Diagn. 2016).
Input DNA may be derived from fresh, fixed, and/or stained cells. Following cell lysis, DNA is digested with a restriction enzyme and adaptors are ligated onto the DNA fragments. Amplification, performed in the same tube, is mediated by a single highly specific PCR primer for all the fragments (approximately 19 million for each haploid genome).
The new fast protocol allows to amplify the whole genome of a single CTC or multiple (up to 20) CTCs in less than 4 hours. The amplified DNA is suitable for library prepration with Ampli1 LowPass kit and Ampli1 OncoSeek panel.
The Ampli1™ QC Kit provides a useful quality control check of the WGA procedure by utilizing a PCR-based assay to establish DNA integrity.
By testing for the presence of one short and three long DNA fragments in the WGA library, the quality of the DNA as starting material for downstream analyses can be assured.
Cells with good quality DNA typically produce 3 or 4 PCR bands, while fixation of cells or cells with degraded DNA will show fewer QC bands. It has been proved that the number of the bands revealed is useful to define a “Genome Integrity Index” (GII) for predicting success of various downstream analytical methods and for assessing the quality of the starting DNA.
Ampli1™ ReAmp/ds Kit is designed to reamplify the Ampli1 WGA product in a reliable and reproducible way. For downstream genetic analysis, you may use the Ampli1 Reamplification product in the same manner as Ampli1 WGA product.
Ampli1 ReAmp/ds Kit can also be used to convert ssDNA molecules into dsDNA molecules both in Ampli1 WGA products and in Ampli1 reamplification products in order to increase the total dsDNA content. This procedure is recommended for accurate DNA quantitation, high-throughput high dimensional downstream analysis, and NGS applications to ensure high sequencing uniformity (but is not required for Ampli1 OncoSeek or Ampli1 LowPass kits).
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The Ampli1 WGA Kit, Ampli1 QC Kit and Ampli1 ReAmp/ds Kit are for Research Use Only. Not for use in diagnostic procedures. Ampli1™ WGA Kits are protected by U.S. patent 6,673,541 and corresponding patent claims outside the United States.