Invitrogen™ Illumina™ TotalPrep™-96 RNA Amplification Kit

Complete system for generating biotinylated, amplified RNA for hybridization with Illumina™ Sentrix™ arrays

Brand:  Invitrogen™ 4393543

Product Code. 10469604

Includes: 110μL T7 Oligo(dT) Primer; 110μL ArrayScript™ Reverse Transcriptase; 220μL 10X First Strand Buffer; 880μL dNTP Mix; 110μL RNase Inhibitor; 110μL RNase H; 1.1mL 10X Second Strand Buffer; 220μL DNA Polymerase; 275μL T7 Enzyme Mix; 275μL Biotin-NTP Mix; 10μL Control RNA; 20mL cDNAPure; 1.1mL RNA Binding Beads; 40mL cDNA Wash Buffer Concentrate; 5.5mL cRNA Binding Buffer Concentrate; 450μL Bead Resuspension Solution; 25mL cRNA Wash Solution Concentrate; 10mL cRNA Elution Buffer; 8 U-Bottom Plates; 8 PCR Plates; 20mL Nuclease-free Water

Description

Using the TotalPrep-96 RNA Amplification Kit
The TotalPrep-96 RNA Amplification Kit incorporates extensive improvements to the in vitro transcription reaction (IVT) over the previous TotalPrep; Kit. The IVT reaction has been optimized for the most effective biotin labeling. An NTP mix containing biotinylated UTP has been added to the kit, allowing ease of use and optimal labeling. These modifications to the kit have significantly increased sensitivity, providing increased Percent Present calls and single-round amplification with just 50ng of input RNA. The kit is based on the RNA amplification protocol developed in the laboratory of James Eberwine (Van Gelder et al. 1990). The procedure consists of reverse transcription with an oligo(dT) primer bearing a T7 promoter using ArrayScript™, a reverse transcriptase (RT) engineered to produce higher yields of first strand cDNA than wild type enzymes. ArrayScript catalyzes the synthesis of virtually full-length cDNA, which is the best way to ensure production of reproducible microarray samples. The cDNA then undergoes second strand synthesis and magnetic-based cleanup to become a template for in vitro transcription with T7 RNA Polymerase. To maximize cRNA yield, MEGAscript™ in vitro transcription (IVT) technology, along with biotin UTP (provided in the kit), is used to generate hundreds to thousands of biotinylated, antisense RNA copies of each mRNA in a sample.

Accessory product
A 96-well magnetic-ring stand is required to use this kit. The 96-well Magnetic-Ring Stand (Cat. No. AM10050) is recommended and designed for paramagnetic bead precipitation from standard 96-well U-bottom microplates. The Magnetic-Ring Stand pellets the magnetic beads in a circle shape, enabling easy removal of supernatant by a robot or multichannel pipettors.

• Perform amplification from just 50ng of input RNA
• Developed for expression profiling using the Sentrix Bead Chips
• Bio-16-UTP now included in optimized NTP mix, reducing set-up time and maximizing labeling
• Includes sufficient reagents for 96 reactions

Expression Array Labeling, Gene Expression Analysis and Genotyping, Microarray Analysis, RNA Amplification

Order Info

Shipping Condition: Approved for shipment on Wet or Dry Ice

Specifications

• Product Type: RNA Amplification Kit
• Includes Label or Dye: Yes
• Reverse Transcriptase: ArrayScript™
• System Type: Illumina Sentrix™
• Product Line: Ambion™, Illumina™ TotalPrep™

• Content And Storage: • 110 μl T7 Oligo(dT) Primer (-20°C)
  • 110 μl ArrayScript™ Reverse Transcriptase (-20°C)
  • 220 μl 10X First Strand Buffer (-20°C)
  • 880 μl dNTP Mix (-20°C)
  • 110 μl RNase Inhibitor (-20°C)
  • 110 μl RNase H (-20°C)
  • 1.1 ml 10X Second Strand Buffer (-20°C)
  • 220 μl DNA Polymerase (-20°C)
  • 275 μl T7 Enzyme Mix (-20°C)
  • 275 μl Biotin-NTP Mix (-20°C)
  • 10 μl Control RNA (-20°C)
  • 20 ml cDNAPure (4°C)
  • 1.1 ml RNA Binding Beads (4°C)
  • 40 ml cDNA Wash Buffer Concentrate (room temperature)
  • 5.5 ml cRNA Binding Buffer Concentrate (room temperature)
  • 450 μl Bead Resuspension Solution (room temperature)
  • 25 ml cRNA Wash Solution Concentrate (room temperature)
  • 10 ml cRNA Elution Buffer (room temperature)
  • 8 U-Bottom Plates (room temperature)
  •
• Labeling Method: Direct Labeling
• Sample Type: Poly(A+) RNA, Total RNA
• Format: Kit
• Quantity: 96 Reactions

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