Invitrogen™ MessageAmp™ Premier RNA Amplification Kit

Innovative kit for the preparation of RNA samples for microarray analysis

Includes: 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

Product Code	Brand	Quantity
10668255	Invitrogen™ 4385821	10 Reactions
10126824	Invitrogen™ AM1792	30 Reactions
10534445	Invitrogen™ 4383452	100 Reactions

Description

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

Order Info

Shipping Condition: Dry Ice

Specifications

• Product Type: RNA Amplification Kit
• Labeling Method: Direct Labeling
• Sample Type: Poly(A+) RNA, Total RNA
• Product Line: Ambion™, MessageAmp™ Premier

• Includes Label or Dye: Yes
• Reverse Transcriptase: ArrayScript™
• Format: Kit

For Research Use Only. Not for use in diagnostic procedures.