The amount of RNA that can be extracted from different biological or clinical samples varies greatly. For example, while a few micrograms of RNA could be easily purified from tissues and cells in excess amounts (such as from a few milligrams of tissue), many liquid biopsy samples may yield very low amounts of RNA. In fact, samples such as urine or plasma may yield 1 – 100 ng or less RNA per 100 µL of sample. Such a range of RNA quantity is often below the detection limit of most commonly used techniques for measuring RNA including nano-spectrophotometry and fluorescent nucleic acid stains. As a result, without properly determined RNA concentration, it becomes very difficult to normalize the starting quantity of RNA used in gene expression studies.
Norgen’s microRNA (cel-miR-39) Spike-In Kit offers a quantified synthetic RNA (cel-miR-39) for spike-in during RNA extraction procedures and subsequent normalization in RT-qPCR assays. The amount of cel-miR-39 RNA recovered after RNA extraction is directly correlated with the amount of total RNA recovered. After reverse transcription (such as with Norgen’s microScript Reverse Transcription system) of the sample RNA (with spike-in), the level of cel-miR-39 can be determined by subjecting the cDNA generated to quantitative PCR (qPCR) using fluorescent nucleic acid stains such as SYBR Green. A cel-miR-39 specific primer is included in the kit. The level of expression of any target transcripts in different RNA samples can now be normalized to the cel-miR-39 transcript level using standard method such as ∆∆Ct relative quantification.
In addition, the cel-miR-39 RNA is compatible to library preparation methods (including ligation-based protocols) in Next Generation Sequencing (Small RNA-Seq) workflows. The cel-miR-39 RNA could be used for normalization as well as for tracking library construction efficiency.
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Storage Conditions
Upon receipt, store Norgen’s microRNA (cel-miR-39) Spike-In Kit at -20°C or lower. Avoid multiple freeze-thaw cycles. If needed, prepare smaller working aliquots and store at -20°C or lower.
| Component | Cat. 59000 |
|---|---|
| cel-miR-39 RNA | 10 pmol |
| cel-miR-39 Forward PCR Primer | 1 nmol |
| Nuclease-Free Water | 1.25 mL |
| Product Insert | 1 |
Norgen’s Total RNA Purification Kits Dx provide rapid methods for the isolation and purification of total RNA from tissue samples, blood, plasma, serum, saliva, bacteria, yeast, fungi and viruses for subsequent in vitro diagnostic use. Extract high quality and purity RNA with excellent RIN values and A260/A280 suitable for downstream applications including qRT-PCR, RT-PCR, microarrays, NGS and more.
Total RNA Purification Kit Dx (Spin Column)
This kit is designed to be used with any downstream application employing enzymatic amplification or other enzymatic modifications of RNA followed by signal detection or amplification. Any diagnostic results generated using the RNA isolated with Norgen’s Total RNA Purification Kit Dx in conjunction with an in vitro diagnostic assay should be interpreted with regard to other clinical or laboratory findings.
Total RNA Purification 96-Well Kits Dx (High Throughput and High Throughput Deep Well)
Thess 96-well kits provide a rapid method for the high-throughput isolation and purification of total RNA in 30 minutes using vacuum manifold, plate centrifuge, or liquid handlers with vacuum capabilities. Total RNA can be isolated from a broad range of sample sources including cultured cells, tissues, blood, serum, plasma, bacteria, yeast, fungi, and viruses.
Intended Use
To minimize irregularities in diagnostic results, suitable controls for downstream applications should be used. Norgen’s Total RNA Purification Kits Dx are intended for use by professional users such as technicians, physicians and biologists experienced and trained in molecular biological techniques. Norgen’s Total RNA Purification Kits Dx do not provide a diagnostic result. It is the sole responsibility of the user to use and validate the kit in conjunction with a downstream in vitro diagnostic assay.
NOTE: This product is not available for sale in the United States.
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| Kit Specifications | |
| Maximum Binding Capacity Per Well | 50 μg |
| Maximum Loading Volume Per Well | 500 μL |
| Size of RNA Purified | All sizes, including small RNA < 200 nt |
| Time to Complete 96 Purifications | 30 minutes |
| Average Yield HeLa Cells (1 x 106 Cells) E. coli (1 x 109 cells) Brain (10 mg) Liver (10 mg) Blood (50 μL, Hamster) | 15 μg 50 μg 9 μg 26 μg 1 μg |
| Maximum Amount of Starting Material: Whole Blood Plasma/Serum Bacteria Yeast Fungi Plant Tissue | 100 µL150 µL1 x 109 cells1 x 108 cells 40 mg 40 mg |
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. These reagents should remain stable for at least 1 year in their unopened containers.
| Component | Cat. Dx17200 (50 preps) | Cat. Dx24300 (192 preps) | Cat. Dx24350 (192 preps) | Cat. Dx24380 (576 preps) |
|---|---|---|---|---|
| Lysis Solution | 40 mL | – | – | – |
| Buffer RL | – | 2 x 40 mL | 2 x 40 mL | 350 mL |
| Wash Solution | 22 mL | – | – | – |
| Wash Solution A | 22 mL | 2 x 38 mL | 2 x 38 mL | 1 x 148 mL 1 x 74 mL |
| Elution Solution | 6 mL | – | – | – |
| Elution Solution A | – | 2 x 20 mL | 2 x 20 mL | 60 mL |
| Mini Spin Columns | 50 | – | – | – |
| 96-Well Isolation Plate | – | 2 | 2 | – |
| 96-Well Isolation Plate (Deep Well) | – | – | – | 6 |
| Collection Tubes | 50 | – | – | – |
| 96-Well Collection Plate | – | 2 | 2 | – |
| 96-Well Collection Plate (Deep Well) | – | – | – | 6 |
| Elution Tubes (1.7 mL) | 50 | – | – | – |
| 96-Well Elution Plate | – | 2 | 2 | – |
| 96-Well Elution Plate (Deep Well) | – | – | – | 6 |
| Adhesive Tape | – | 4 | 4 | 12 |
| Product Insert | 1 | 1 | 1 | 1 |
