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Top Five Ways to Accelerate Your Research by Combining Ambion & Applied Biosystems Products
Together, Ambion, an Applied Biosystems Business, and Applied Biosystems provide a complete family of products for sample preparation through target detection and analysis. RNA interference (RNAi) research, microRNA (miRNA) research, and gene expression analysis are major applications where the combination of Ambion and Applied Biosystems product lines work together to create optimized workflow solutions.
1. Enhancing RNAi Success
Silencer® siRNAs
MagMAX™-96 Total RNA Isolation Kit
TaqMan® Gene Expression Assays
TaqMan® Universal PCR Master Mix
7900HT Fast or 7500 Real-time PCR Systems
RNA interference (RNAi) enables scientists to elucidate gene function in a matter of days; the correct protocol and optimized reagents are essential. Applied Biosystems and Ambion provide the most complete suite of validated, quality reagents for RNAi experiments. Our potent Silencer siRNAs, optimized TaqMan Gene Expression Assays, TaqMan Universal Master Mix, and Real-Time PCR Systems, along with the supplied protocols, streamline each step of an RNAi experiment and remove the technical variability from your experiments, enabling you to focus on making critical biological discoveries.
Gene silencing experiments require siRNAs that efficiently knock down the target of interest. To prove that a particular siRNA sequence is effective, the siRNA should be functionally tested in cells and validated to reduce target mRNA levels by a certain amount. Silencer siRNAs eliminate the guesswork and tedious labwork associated with siRNA design and testing. Silencer Pre-designed siRNAs are designed with a proprietary algorithm and are guaranteed to silence your target gene, while the Silencer Validated siRNAs have also been shown experimentally to knock down target gene expression by at least 70% as measured by TaqMan Gene Expression Assays (data provided with purchase).
Once siRNAs are delivered, RNA can be isolated from the transfected cells using MagMAX Total RNA Isolation Kits, in either manual or automated high throughput mode, yielding RNA ideal for subsequent real-time PCR analysis.
Real-time PCR is the preferred method for evaluating an siRNA's knockdown efficiency. Gene-specific, ready-to-run TaqMan Gene Expression Assays, which are pre-designed primer:probe sets for real-time PCR, provide a rapid and simple way to measure siRNA-induced target knockdown.
Find Silencer siRNAs and corresponding TaqMan Gene Expression Assays for your genes of interest.
2. MicroRNA Research Made Easy
mirVana™ miRNA Isolation Kit
TaqMan® MicroRNA Reverse Transcription Kit
TaqMan® MicroRNA Assays
7900HT Fast or 7500 Real-Time PCR Systems
MicroRNAs (miRNAs) are small RNA molecules that regulate gene expression through effects on translation or stability of target RNAs. More than one third of human genes may be regulated by miRNAs, and each miRNA is believed to regulate multiple genes. To understand miRNA function, scientists must determine which miRNAs are expressed, where and when they are expressed, the cell processes that they regulate, and the genes that they control. However, studying miRNAs can be a challenge because of their small size.
Start your miRNA study on solid footing by isolating total RNA containing small RNAs from your samples using the mirVana miRNA Isolation Kit. Then rely on the Applied Biosystems validated, ready-to-use system for quantitation of miRNA expression levels—TaqMan MicroRNA Assays run on the 7900HT Fast Real-Time PCR System.
TaqMan MicroRNA Assays are the most accurate method for evaluating miRNA expression levels. Orginally they were developed for quantitating expression of one to several different microRNAs; however, by creating ~48 miRNA-specific cDNAs in a single reaction using the MicroRNA Multiplex-RT Pools, the TaqMan RT-PCR approach rivals array hybridization as a microRNA expression profiling tool. With a dynamic range of 6-7 logs compared to only 2-3 for microarrays, results in ~3 hours compared to 1-2 days, and minimal RNA sample requirements (1-10 ng), the benefits of TaqMan Assays over microarrays are significant for miRNA quantitation. The Applied Biosystems validated miRNA expression profiling platform will accelerate discovery of the many biological roles of miRNAs in cells and in human diseases. For more information on TaqMan miRNA products, visit mirna.appliedbiosystems.com. For more information about the entire suite of Ambion miRNA analysis tools, visit the miRNA Resource.
3. MicroRNA Expression Profiling and Validation Using Array Analysis and Quantitative RT-PCR
mirVana™ miRNA Probe Set V2
TaqMan® MicroRNA Assays
7900HT Fast or 7500 Real-Time PCR Systems
Microarray analysis is the most commonly used technique for microRNA expression profiling; the Ambion mirVana miRNA Probe Set V2 is a collection of amine-modified DNA oligonucleotide probes targeting human, mouse, and rat miRNAs from the miRBase Sequence Database. The mirVana miRNA Probe Set V2 also includes probes targeting an exclusive set of 152 novel human miRNAs called Ambi-miRs that were identified through a combination of bioinformatic prediction, cloning, and detection in human total RNA samples. These sequences are specifically designed for the production of glass slide miRNA arrays.
As with any array hybridization approach to expression profiling, array results require validation. Applied Biosystems TaqMan MicroRNA Assays offer a simple, direct, and rapid way to validate microRNA expression profiling results obtained using the mirVana miRNA Probe Set. Compared to other methods such as Northern analysis and nuclease protection strategies, TaqMan MicroRNA Assays are quicker, easier, and more robust. Get answers about miRNA expression using mirVana miRNA Bioarrays and validate your results using TaqMan MicroRNA Assays on Applied Biosystems Real-Time PCR Systems.
4. Gene Expression Analysis Directly from Cell Lysates
Cells-to-cDNA™ II Cell Lysis Buffer
High Capacity cDNA Reverse Transcription Kit
TaqMan® Gene Expression Master Mix
TaqMan® Gene Expression Assays
7900HT Fast Real-Time PCR System
Traditionally, the first step in gene expression experiments has been to recover pure RNA from experimental samples. Even using the quickest techniques for isolating RNA requires a significant amount of hands-on manipulation and time. Applied Biosystems and Ambion now offer a robust solution for capturing gene expression information while skipping RNA isolation from cultured cells. The combination of a simplified sample preparation step from the Cells-to-cDNA technology, combined with the sensitivity and specificity of TaqMan chemistry, enables real-time PCR directly from cell lysates. The innovative reagent, Cells-to-cDNA II Cell Lysis Buffer, lyses cultured mammalian cells (1 x 102 to 1 x 106 cells) while protecting endogenous RNA from degradation. This lysate can serve as an effective template for real-time RT-PCR without having to first isolate the RNA. Cells-to-cDNA II lysates can be efficiently reverse transcribed using the High Capacity cDNA Reverse Transcription Kit containing MMLV. Then, expression levels can be quantitatively detected using the TaqMan Gene Expression Master Mix, state-of-the-art TaqMan Gene Expression Assays, and trusted Real-Time PCR Instruments from Applied Biosystems.
5. Reliable SNP Genotyping Analysis from Formalin-fixed, Paraffin-embedded (FFPE) Tissue
RecoverAll™ Total Nucleic Acid Isolation Kit for FFPE
TaqMan® RNase P Detection Reagents (FAM™ Dye)
TaqMan® SNP Genotyping Assays
7900HT Fast Real-Time PCR System
The ability to isolate nucleic acids that are suitable for molecular analysis from archived formalin-fixed, paraffin-embedded (FFPE) tissue, enables the comparison of healthy and diseased tissues at both the genomic and gene expression levels through the use of techniques like single nucleotide polymorphism (SNP) genotyping and real-time RT-PCR. The combined use of optimized reagents and protocols from Ambion and Applied Biosystems produces reliable and reproducible SNP genotyping data from FFPE samples. Isolate DNA from FFPE samples using Ambion’s RecoverAll Total Nucleic Acid Isolation Kit, accurately detect percent functional DNA in samples using TaqMan RNase P Detection Reagents (FAM), and obtain consistent genotyping results using TaqMan SNP Genotyping Assays.
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