A technical poster from Fluidigm Europe that demonstrates a new approach, which when used in combination with Fluidigm’s BioMark real-time PCR system, provides a simple, inexpensive and reproducible solution to single cell gene expression.
The authors of the poster demonstrate the use of the Fluidigm 48.48 Dynamic Array integrated fluidic circuit (IFC), as well as a technique known as specific target amplification (STA). Each 48.48 Dynamic Array is shown to systematically combine samples and assays into 2,304 real-time qPCR reactions, of final reaction volumes of 6.75 nanolitres. Experimental results are shown using FACS-sorted single cells from a human colorectal carcinoma cell line (HCT 116).
Using the Fluidigm 48.48 Dynamic Array IFC, the researchers were able to conduct gene expression studies from single cells, with excellent sensitivity. While significant differences in expression levels of the same genes across multiple (supposedly identical) single cells was observed, the BioMark / 48.48 Dynamic Array platform is shown to produce extremely consistent results when identical amounts of actual transcript were tested. A copy of the technical poster may be downloaded from www.fluidigm.com/pdf/posters/FLDM_BMK_MRKT00098.pdf.
The Fluidigm 48.48 Dynamic Array IFC provides a highly efficient solution for single cell gene expression quantification studies using off-the-shelf TaqMan PCR assays and reagents. The key to 48.48 Dynamic Array is the matrix of channels, chambers, and valves that do the work of assembling assays. Using the product – scientists can enjoy the same qPCR data quality and experiment flexibility that they are currently receiving from a 384-well microplate system while increasing throughput 24X. To run an experiment – 48 samples and 48 assays are loaded into the inlets of the Dynamic Array input frame and pressure loaded into 2,304 reaction chambers. Using this simple approach scientists are able to test 48 genes against 48 samples in a single run. The savings in time and effort add up rapidly. For example, a 2,000-sample study that could be completed with 4,032 pipetting steps using the Fluidigm 48.48 Dynamic Array would require 192,000 steps to complete the same study using microplates. With dynamic arrays, genetic researchers can advance their research much more easily and affordably, and they can do so while using gold-standard TaqMan assays.
Filed Under: Drug Discovery
