EmbryoCellect™ uses microarray technology (array Comparative Genomic Hybridisation or aCGH) to compare the number of chromosomes in a sample cell to a known reference sample. The samples are labelled with different fluorescent dyes and the amount of fluorescence is measured and compared for each chromosome.
This provides a reliable and straightforward way to count the number of chromosomes in the test cell and detect whole chromosome aneuploidy.
EmbryoCellect™ is for research use only and should not be used as a clinical diagnostic.
Samples can be processed in batches as small as four, which makes the workflow more flexible.
The EmbryoCellect™ kit contents
- Cell lysis buffer and enzyme for lysing the sample cells
- Reference male gDNA
- Whole Genome Amplification (WGA) reagents
- Fluorescent labelling PCR reagents
- Five patented EmbryoCellect™ microarray slides with four microarrays per slide
- Software: A Simple Excel macro that uses a microarray scanner-generated data file
The EmbryoCellect™ Microarray workflow
|Cell lysis||Following biopsy, a gentle but effective enzyme-based lysis procedure ensures robust cell lysis and a readily accessible DNA template for whole genome amplification – 15 mins|
|Whole genome amplification||Whole genome amplification is performed using RHS’ DOP-PCR, which has been optimised for the RHS microarray. DOP-PCR uses degenerate primers to initiate DNA amplification, binding across a broad range of different sequences scattered genome wide -2 ½ hours|
|Agarose gel assessment||Following amplification, the use of agarose gel electrophoresis is recommended to ensure that cell amplification has been successful – 30 mins|
|Labelling PCR||Successfully amplified samples are fluorescently labelled by a second DOP-PCR. The test is labelled with a Cy3 equivalent dye and the reference with a Cy5 equivalent dye – 45 mins|
|Clean-up and nanodrop Agarose gel assessment||Once purified, these labelled amplicons are again assessed using agarose gel electrophoresis and spectrophotometry to ensure adequate amplification and dye incorporation has occurred – 1 hour|
Microarray washing, Microarray scanning and analysis
|Samples are competitively hybridized to the RHS microarray – 3 hours, to overnight
After incubation, the microarray is washed and scanned (1 hour). The ratio of test to reference dye intensity after normalization is determined using RHS proprietary analysis method, providing the ploidy status of each chromosome in each sample.