Introducing new genes into your cells, or silencing others, is easier each year as the technology and reagents for cellular have steadily improved. And the improvement shows...
The entire foundation of modern molecular biology is predicated on a single supposition: That it is possible to deliver nucleic acids to cells. From reporter gene assays to RNAi, knockout mice to...
Mary Kay Bates discusses electroporation of nucleic acids to cultured cells using the Ingenio electroporation solution.
Certain cell-lines of immune or neurological origin, as well as primary cells, are traditionally considered 'hard to transfect' cells, as these are resistant to chemical transfection reagents. Electroporation is a physical means of transfection that enables delivery of nucleic acid into cells by electropermeabilization and electro-transfer. Electroporation is an essential tool for nucleic acid delivery that can be applied to a wide variety of cell types, especially hard to transfect cells. Selection of electroporation solution and optimization of pulse parameters are critical steps to successful electropermeabilization using any electroporation device.
Cancer diagnostics just ain't what they used to be – and that's a good thing. Classic tests like mammograms, digital-rectal exams, and prostate-specific antigen tests, are being joined by a new class of molecular diagnostics based on nucleic acid detection.
Despite the amazing ability of siRNAs to silence gene expression in relatively straightforward experiments — at least compared to creating a knockout mouse — there are still a few wrinkles to iron out when it comes to transfecting them. Small (or short) inhibitory (or interfering) RNAs are small pieces of RNA that enter cells and interfere with the expression of their target gene, thereby silencing its expression. However, introducing siRNAs into cells is still proving a challenge — one thatwhich has future implications for using siRNAs therapeutically.
Finding the right transfection reagent is a key step in your cell-based expression studies. You need a reagent that will effectively deliver your construct without causing cellular changes that could interfere with the interpretation of your results. In addition to these essentials, it’d be nice...
MicroRNA (miRNA) expression profiling is commonly used to assess the relative levels of individual miRNAs in specific tissues. To achieve accurate results using microarray detection strategies, it is important that all sample miRNA species are representatively labeled prior to the hybridization step. In this study, we compared the expression profiles of miRNAs labeled using enzymatic and chemical methods. Using miRNA samples isolated from mouse brain and heart, we show that some specific miRNAs are not detected when enzymatic tailing methods are used; however they are detected with the direct chemical labeling method (Label IT®). The presence of the discrepant miRNAs in the model mouse tissues was validated by northern blot analysis and qRT-PCR. Thus, miRNA expression profiles generated by enzymatic labeling methods (poly(A) polymerase tailing) may not represent all miRNAs present in a given sample. The Label IT technology offers a more sensitive and universal miRNA labeling method.
RNA interference (RNAi) is a promising new gene silencing technology of great use in studying the functions of genes and understanding the genetic basis of cellular physiology in mammals. This inexpensive silencing of target gene expression enables target repression of individual genes without having to resort to costly knockout models. TransIT®-siQUEST™ Transfection Reagent is a highly efficient...
One way for researchers to deduce a gene products’ function is to eliminate that gene, hence eliminating the protein, in an organism and observe the resulting phenotype. Traditionally this has been a complicated process, but recent innovations in products for RNA interference (RNAi) have made it much simpler...