Description
Delivering High Quality gDNA from a Single Cell Genomic analysis from single cells is an emerging area as researchers focus in on differences within cell populations. The amount of genomic DNA needed to carry out a full range of testing has impacted progress, and while techniques become more sensitive, sharing a single cell's DNA across platforms is not possible. Single Cell GenomiPhi DNA Amplification kit has been optimized to wholly amplify genomic DNA from as little as a single cell, generating micrograms of high quality DNA for use in downstream applications. At very low levels of input DNA, successful amplification relies on reagent quality. Single Cell GenomiPhi has been developed with this in mind. Lysis reagents are optimized to fully release genomic DNA from the cell and subsequently denature the DNA to enable optimal amplification and coverage. Background amplification, often associated with Multiple Displacement Amplification (MDA), is suppressed throughout the incubation so that only input DNA is amplified. High quality reagents and an optional, proprietary enzymatic clean-up step in the protocol ensure that any potential DNA contaminants introduced during set-up are removed before each individual reaction. In addition, our manufacturing processes, including UV and enzymatic reagent cleanup, help to ensure that all kits are free from any detectable DNA contamination and enable sensitivity of amplification down to 1 fg of gDNA. Femtograms to Micrograms using a Simple Workflow Single Cell GenomiPhi offers a simple two-step workflow for whole genome amplification. The amplification process is 2 to 4 hours depending on the amount of starting DNA and yields of 4 to 7 µg can be achieved. Amplification is highly uniform over the entire genome so that locus representation remains extremely close to the original DNA sample. Furthermore, amplification is carried out with very high fidelity due to Phi29 DNA polymerase proofreading activity. DNA from single cells can be reproducibly and reliably amplified for use in many downstream applications. Single Cell GenomiPhi has been successfully used to generate template for SNP array genotyping, array CGH, and next-generation sequencing.
