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Ampli1 Whole Genome Amplification

Ampli1 Whole Genome Amplification

Balanced and robust whole genome amplification for reliable single cell analysis

Successful analysis of single cell genomes starts with an appropriate Whole Genome Amplification (WGA) step that provides a robust and reproducible amplification efficiency for every locus and on each allele.

Ampli1™ WGA method is the only fully “non-random” whole genome amplification method for single, human cells. It provides a robust and reproducible amplification efficiency achieving the lowest Allelic Drop Out (ADO) rate compared to other methods1-6 so you don’t miss out on key genetic data.

The new Ampli1 WGA PLUS Kit has been optimized to perform whole genome amplification in less than 4 hours for all compatible sample types (live cells, fixed or fixed and permeabilized cells) and is suitable for high throughput liquid handling automation.
 

 

Reliable, reproducible, balanced single cell amplification

Key features of the Ampli1 WGA PLUS Kit:

●    Robust and reproducible protocol produces a library of highly concentrated DNA to run multiple downstream analyses. Single-tube, no-precipitation procedure prevents template loss;

●    Single, highly-specific, primer-mediated PCR ensures balanced amplification of both alleles achieving the lowest Allelic Drop Out rate;

●    Compatible with automatic liquid handlers (such as Hamilton Starlet); Fast protocol performs whole genome amplification in 4 hours, with less than 1 hour hands-on time;

●    Double-stranded WGA products (no-need for additional double-stranding reactions for capture-based, targeted-sequencing downstream applications);

●    No chimeric artifacts and low amplification noise avoids calling of false structural variants and produces reliable and clean copy number profiles;

●    Broader genome coverage with higher uniformity.

Ampli1 WGA PLUS Kit

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 and Ion Torrent, allow for the detection of hotspot mutations-focal copy number alterations and genome-wide copy number alterations respectively, starting from the same single cell. 

The product is for research use only (RUO) – not for use in diagnostic procedures.
 

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Ampli1 WGA QC Kit

The Ampli1 WGA QC Kit is a multiplex PCR-based assay providing a useful quality control check of the WGA procedure.

By testing for the presence of one short and three long DNA fragments in the WGA library, the input DNA integrity and suitability of the WGA amplification product for further downstream analyses can be assessed.

Cells with good quality DNA typically produce 3 or 4 PCR bands, while fixed cells or cells with degraded DNA will show fewer QC bands. The number of bands 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.

Product is for research use only (RUO) – not for use in diagnostic procedures.

References

1Binder V et al, Hum Mutat. 2014 Oct;35(10):1260-70.

2Huang L et al, Annu Rev Genomics Hum Genet. 2015;16:79-102.

3Babayan A et al, Oncotarget. 2017; 8:56066-56080.

4Borgström E et al, PLoS ONE 2017 12(2): e0171566.

5Normand E et al, Prenat Diagn. 2016 36(9): 823-830.

6Raz O et al, Int J Mol Sci. 2022 23(11):6161. 

Products Resources

Visit our product resources page to access Instructions for Use and Material Safety Data Sheets (MSDSs) for all our products.

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Discover recently published articles featuring research enabled by our CELLSEARCH, DEPArray and Ampli1 technologies

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