Good-quality RNA impacts downstream analysis and results, explains Lucy Sha, marketing director for gene and protein discovery at GE Healthcare (www.gehealthcare.com). “Sample preparation is critical, and the entire process of RNA analysis should be seamless, without bottlenecks,“ Sha adds.
William Dietrich, product manager for RNA analysis at Stratagene (www.stratagene.com), agrees, adding, “Trends in RNA analysis include getting value from a product—saving time and money and simplifying the workload and process—by getting complete solutions from one supplier.“
At GE Healthcare, “we´ve taken the products we´ve created for nucleic acid amplification and purification and brought the same expertise to sample preparation,“ Sha says. “The objective is to let researchers focus more time and energy on the creative aspects of their work.“
Total RNA isolated using the GE Healthcare RNAspin Isolation kits is consistently of high quality and good yield, Sha notes. The quality is sufficient for use in transcription profiling workflows like microarray analysis, which takes the total RNA through cDNA synthesis and labeled-cRNA amplification to loading on microarray and expression analysis.
cRNA generated using this total RNA is also reproducibly of high quality, according to the company. Individual total RNA samples are highly uniform in their quality and transcript population representation, which is crucial for microarray experiments.
Sample preparation for RNA isolation and gene-expression measurement is a key area for Stratagene. One of its products enables gene-expression researchers to lyse and stabilize cells and go directly to the RT-PCR process.
Stratagene´s SideStep cDNA synthesis and QRT-PCR products that use the company´s Brilliant QRT-PCR technology are designed to deliver efficient cell lysis, RNA stabilization, and sensitive quantitative gene expression analysis without RNA purification. Enabling researchers to screen multiple samples first, they need only purify the most relevant, interesting RNA samples, saving over 70% of the cost and 60% of the time it takes to column purify RNA, according to Dietrich.
SideStep lysis and stabilization technology stabilizes RNA for up to six months using a single-tube format, minimizing RNA loss and degradation. The easy-to-use, nontoxic method accommodates a wide range of cell input, making it a flexible method for immediate gene-expression measurement from cells.
Applications include detecting gene expression of siRNA-induced knockdown experiments, gene-expression measurement in real time using SYBR or probe-based chemistry, miRNA detection, low-abundance message detection, mRNA enrichment directly from cells, producing high-quality, full-length QPCR-grade cDNA directly from cells, and analyzing laser microdissected cells from fresh or frozen tissues.