Isolation of genomic DNA from formalin fixed paraffin embedded (FFPE) tissues is a critical step for molecular diagnostic (MDx) assays. Here we investigate extraction of DNA from FFPE using five comparable methods.
After all the hard work of editing your cell line, you want to have confidence in your new research model. So, how do you verify your cell line is what you expect it to be? Could a heterogeneous cell population be obscuring your editing effects? Is observed phenotype being caused by the targeted gene edit, or unintended off-target effects? Here we discus ways to add supporting data to validate your gene-engineering projects.
CRISPR technology now allows genes and molecular pathways to be examined with greater definition. We look at how knockout cell lines, either together with gene rescue and replication of disease mutations or as an independent cell model, can be used to validate your research and extend your findings
With new advances in technology, we're now moving more towards large panel next-generation sequencing (NGS) assays and whole exome sequencing (WES). NGS is gaining popularity thanks to high-throughput capability and reduction in cost per sample.
Essential genes are defined as genes that are critical for the survival of an organism. These are considered to be genes that are absolutely required for the cell to grown, proliferate and survive. Deletion of an essential gene from a cell eventually leads to the death of this cell or a severe proliferation defect. As a consequence, it is impossible to generate cells with a knock-out or deletion of essential genes.
Thanks to next-generation sequencing (NGS), we are starting to understand the mutational changes that occur across the board in the cancer genome. With this knowledge comes potential - novel mutated genes and the proteins that they encode are candidates for prognostic markers and/or new drug targets.