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™ 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.

• Detect total RNA (including miRNA) from PAXgene™ 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.

Simple Workflow Without Sacrificing Sensitivity & Specificity
The process consists of pelleting and washing a 500uL 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. 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’s 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
Reverse transcription reagents for cDNA synthesis, TaqMan™ master mixes, and TaqMan™ assays for real-time PCR analysis are sold separately and are not included with the kit.

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™ 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. Our 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 = 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.

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 (including miRNA) from PAXgene™ Blood RNA Tubes using proven magnetic bead-based separation technology.

One-Stop Shopping: The Stabilized Blood-to-C_T™ Nucleic Acid Preparation Kit for qPCR is compatible with our suite of products (including RT and PCR master mixes that must be purchased separately). Simplify the ordering process and reduce freight cost by getting more from one supplier.

Specifications:
• Compatibility— PAXgene™ Blood RNA Tubes
• Processing Time—Under 1 hour
• Format— microfuge tubes, 96-well plates
• Equipment Needed—Standard centrifuge capable of 5,000 x g with either 96-well plate or microfuge tube adaptors and 96-well plate or microfuge tube vortexer
• Number of Preps —200 reactions

NOTE: Blood tubes, reverse transcription reagents for cDNA synthesis, and TaqMan™ master mixes for real-time PCR analysis are sold separately.

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

PAXgene™ and PreAnalytiX™ are registered trademarks of PreAnalytiX GmbH. No sponsorship, endorsement, or affiliation is implied herein.