Invitrogen™ Stabilized Blood-to-C_T™ Nucleic Acid Preparation Kit for qPCR, compatible with either PAXgene™ or Tempus™ Blood RNA Tubes

The Stabilized Blood-to-C_T™ Nucleic Acid Preparation Kit for qPCR makes it possible to perform expression analysis directly from a 500 μL aliquot of human whole blood collected in PAXgene™ or Tempus™ Blood RNA tubes without conventional RNA purification.

Supplier:  Invitrogen™ 4449079

Catalog No. 44-490-79

Includes: Kit Components: Box 1 - Digestion Solution 5mL, Tempus™ Pellet Enhancer 1mL, Tempus™ Wash No. 1 37.5mL, Tempus™ Wash No. 2 37.5mL, 2X PBS for Tempus™ 12.5mL, PAXgene™ Wash 75mL. Box 2 - Stop Solution 0.5, DNase 1, 50μL

Description

The Stabilized Blood-to-C_T™ Nucleic Acid Preparation Kit for qPCR makes it possible to perform expression analysis directly from a 500 μL aliquot of human whole blood collected in PAXgene™ or Tempus™ Blood RNA tubes without conventional RNA purification. This simple procedure creates a lysate that can go directly into reverse transcription and real-time PCR with the sensitivity and specificity equivalent to conventionally purified RNA. This kit saves time, generates much less waste, and offers a simple workflow that is suitable for a few samples or can easily be incorporated into automated, high throughput applications.

Simple Workflow Without Sacrificing Sensitivity & Specificity
The process consists of pelleting and washing a 500μL aliquot from the blood tube, followed by a unique digestion/DNase step which preserves RNA integrity. The preparation takes place entirely at room temperature unlike other products that require temperature-specific incubation steps. Up to 96 samples can be prepared in less than an hour, half the time it takes for competitive spin column purification to process only 12 samples, with less hands-on effort. The resulting lysate can go directly into one-step or two-step qRT-PCR reactions or stored for later use (1 hour at 4°C or 5 months frozen). This straightforward procedure takes place in the same sample processing well, from start to finish, meaning no sample transfers. Reverse transcription reagents for cDNA synthesis, TaqMan master mixes, and TaqMan assays for real-time PCR analysis available separately.

The total number of reverse transcription and PCR reactions possible from one starting sample depends on the minimum sample requirements of the desired RT-PCR kits. Please see the user documentation for more details.

Screen, Discover, or Validate
For users with specific genes of interest, the Stabilized Blood-to-C_T Nucleic Acid Preparation Kit for qPCR was designed to enable quick screening from a small amount of starting sample (approximately 5% of the total volume of a properly filled PAXgene or Tempus Blood RNA Tube per sample). The remainder of the blood tube can be archived for later use or purified as needed. Those who perform discovery or validation experiments with real-time RT-PCR analysis will save effort, time, and overall cost with the Stabilized Blood-to-C_T Nucleic Acid Preparation Kit for qPCR – while employing a scalable process allowing labs bottlenecked by conventional purification to finally start processing samples with ease.

miRNA Analysis
With the growing focus on non-coding RNA and its regulatory role in development and disease, it's reassuring to know these small but important RNA molecules can be detected properly with the Stabilized Blood-to-C_T Nucleic Acid Preparation Kit for qPCR. Scientists have tested the lysate with the Applied Biosystems TaqMan MicroRNA Reverse Transcription Kit and TaqMan Universal Master Mix II against a large panel of TaqMan microRNA assays and generated results equal to or better than those of the leading competitor's miRNA purification product.

Reduced Footprint & Less Waste for More Savings
Competitive products, especially column-based methods, generate considerable amounts of plastic and biohazardous waste per sample (spin columns, tips, microfuge tubes, and reagents). The Stabilized Blood-to-C_T Nucleic Acid Preparation Kit for qPCR simplifies the workflow so the total plastic waste is 75% less by weight for a comparable number of samples processed via columns. The kit generates no organic waste. It also has a space-saving footprint to maximize freezer capacity. Reduced hazardous waste disposal and less plastic consumption translate into cost savings – as well as reducing the environmental impact.

• Detect total RNA from PAXgene (including miRNA) or Tempus Blood RNA Tubes without purification
• Fast—Prepare 96 samples for TaqMan-based real-time RT-PCR in under 1 hour
• Robust—500 μl sample processed at room temperature with results equivalent to those from purified RNA
• Flexible Format—Samples can be prepared in microfuge tubes or manual/automated 96-well plates
• Compatible with 1-Step, 2-step qRT-PCR and automated protocols, high-throughput compatible

The real-time RT-PCR data generated from the lysate is equivalent to results obtained from conventionally purified RNA from the same tubes and has been validated with the following qPCR reagents:

• Applied Biosystems High Capacity RNA-to-cDNA Kit and TaqMan Gene Expression Master Mix
• Invitrogen SuperScript™ VILO™ cDNA Synthesis Kit and TaqMan Gene Expression Master Mix
• Applied Biosystems TaqMan MicroRNA Reverse Transcription Kit and TaqMan Universal Master Mix II
• Applied Biosystems TaqMan One-Step RT-PCR Master Mix Reagents Kit

For Users Needing Full Purification:If conventional RNA purification is still required for the remnant blood tube, one option is the MagMAX for Stabilized Blood Tubes RNA Isolation Kit. This product maximizes your yield of total RNA from PAXgene (including miRNA) or Tempus Blood RNA Tubes using proven magnetic bead-based separation technology.

Specifications

• Content And Storage:

  Kit Components:
  Box 1-Store at 4°C:
  • Digestion Solution, 5 mL
  • Tempus™ Pellet Enhancer, 1 mL
  • Tempus™ Wash #1, 37.5 mL
  • Tempus™ Wash #2, 37.5 mL
  • 2X PBS for Tempus™, 12.5 mL
  • PAXgene™ Wash, 75 mL
  
  Box 2-Store at -20 °C
  • Stop Solution, 0.5 mL
  • DNase 1, 50 μL

• Purification Time: <1 hr.
• Final Product Type: Lysate
• For Use With (Application): 1-Step qRT-PCR, 2-Step qRT-PCR
• No. of Reactions: 50 Preps
• Quantity: 50 preps

• Format: Tube, 96-well Plate
• Sample Type: Blood
• High-throughput Compatibility: High-throughput Compatible, Not High-throughput Compatible (Manual), Automated Protocols
• Green Features: Fewer resources used and less waste, Less hazardous
• Product Line: Ambion™, Blood-to-CT™
• Target: Lysate

Safety and Handling

WARNING: Cancer and Reproductive Harm - www.P65Warnings.ca.gov

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