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Invitrogen™ MessageAmp™ Premier RNA Amplification Kit
Beskrivning
Innehåller
110μL First Strand Enzyme Mix; 440μL First Strand Buffer Mix; 220μL Second Strand Enzyme Mix; 550μL Second Strand Buffer Mix; 660μL T7 Enzyme Mix); 2 x 1.1mL T7 Biotin IVT Mix; 10μL Control RNA 1mg/mL; 1.75mL Nuclease-free Water; 6mL aRNA Binding Buffer Concentrate; 1.2mL RNA Binding Beads; 30mL aRNA Wash Solution Concentrate; 15mL aRNA Elution Solution; 1mL 5X Array Fragmentation Buffer; 2 U-Bottom Plates 4 PCR Plates; 1 Reservoir; 10mL Nuclease-free Water
Kit employs the proven methodology for RNA sample preparation and labeling based on the original T7 in vitro transcription (IVT) amplification technology, known as the Eberwine or reverse transcription-IVT (RT-IVT) method. The RT-IVT method is considered the gold standard for sample preparation in microarray-based expression profiling. It is well documented in the current scientific literature and was experimentally validated using TaqMan™ RT-PCR. Builds upon the linear RNA amplification method and reagents developed for MessageAmp™ II Kits with enhancements that create a simplified workflow, improved performance, and more flexible RNA input requirements.
Using the MessageAmp Premier RNA Amplification Kit
Based on this proven enzymatic amplification and labeling strategy, the MessageAmp Premier procedure starts with a simple cDNA synthesis reaction using ArrayScript™ reverse transcriptase. No cleanup step is required after the RT; cDNA is used directly in a high yield IVT reaction using MEGAscript technology. The IVT is configured to incorporate the modified nucleotide, biotin-UTP, into the aRNA synthesized. (In this protocol, the amplified RNA is referred to as aRNA; in the literature, it is also commonly called cRNA.) Once purified, the aRNA is suitable for use on microarray gene expression analysis platforms designed for biotin-modified antisense RNA samples.
- Very flexible RNA input requirements: 20-500ng total RNA, depending on the tissue or cell type
- Streamlined workflow: depending on the RNA input amounts, only a single day of processing time is required to complete a microarray experiment
- Simplified protocol and streamlined reagents
- Complete kit: everything needed to prepare biotin-modified aRNA is included
- Rigorous quality control that includes GeneChip™ analysis
- T7 MEGAscript™ IVT delivers up to 50,000-fold amplification
- Single-tube format
- Easy-to-use, high-recovery, magnetic-bead aRNA purification
Lowest RNA input requirements for a single round of amplification:
- 20ng total RNA yields enough biotin-modified aRNA for most microarray formats, with results equivalent to those from samples prepared from significantly more starting material using other RT-IVT kits
Expression Array Labeling, Gene Expression Analysis and Genotyping, Microarray Analysis, RNA Amplification
Beställningsinfo
Shipping Condition: Dry Ice
Specifikationer
Specifikationer
| Produkttyp | RNA Amplification Kit |
| Innehåll och lagring | • 11 μl First Strand Enzyme Mix (-20°C) • 44 μl First Strand Buffer Mix (-20°C) • 22 μl Second Strand Enzyme Mix (-20°C) • 55 μl Second Strand Buffer Mix (-20°C) • 66 μl T7 Enzyme Mix (-20°C) • 220 μl T7 Biotin IVT Mix (-20°C) • 10 μl Control RNA 1 mg/ml (-20°C) • 1.75 ml Nuclease-free Water (any temperature) • 600 μl aRNA Binding Buffer Concentrate (room temperature) • 120 μl RNA Binding Beads (4°C) • 10 ml aRNA Wash Solution Concentrate (room temperature) • 5 ml aRNA Elution Solution (4°C or room temperature) • 1 ml 5X Array Fragmentation Buffer (room temperature) • 10 8-Strip PCR Tubes & Caps (room temperature) • 1 U-Bottom Plate (room temperature) • 1 Reservoir (room temperature) • 10 mL Nuclease-free Water (any temperature) |
| Inkluderar etikett eller färgämne | Yes |
| Märkningsmetod | Direct Labeling |
| Omvänt transkriptas | ArrayScript™ |
| Provtyp | Poly(A+) RNA, Total RNA |
| Formatera | Kit |
| Produktlinje | Ambion™, MessageAmp™ Premier |
| Kvantitet | 10 Reactions |
For Research Use Only. Not for use in diagnostic procedures.
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