This kit provides a rapid method for the isolation and purification of small RNA molecules (< 200 nt) from cultured animal cells, small tissue samples, bacterial cells, plants, and blood.
Two columns are provided with this kit. The first column captures large RNA, while the small RNA are captured on a second column and are eluted concentrated in 25 µL of nuclease-free water.
The small RNA can be used in various downstream applications relating to miRNA profiling, gene regulation, and functional analysis. The eluted RNA is ready for RT-qPCR, microarrays, and NGS applications.
Background
These small RNAs include regulatory RNA molecules such as microRNA (miRNA) and short interfering RNA (siRNA), as well as tRNA and 5S rRNA. Small RNA molecules are often studied due to their ability to regulate gene expression. miRNAs and siRNAs are typically 20-25 nucleotides long and regulate gene expression by binding to mRNA molecules and affecting their stability or translation.
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| Kit Specifications | |
| Maximum Column Binding Capacity | Up to 50 μg RNA |
| Maximum Column Loading Volume | 650 μL |
| Minimum Elution Volume | 20 μL |
| Size of RNA Purified | All sizes, including small RNA < 200 nt |
| Time to Complete 10 Purifications | 25 minutes |
| Maximum Amount of Starting Material: Animal Cells Animal Tissues Bacteria Plant Tissues Blood | 3 x 106 cells 5-25 mg 1 x 109 cells 50 mg 100 μL |
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.
| Component | Cat. 21300 (25 preps) |
|---|---|
| Buffer RL | 40 mL |
| Wash Solution A | 38 mL |
| Elution Solution A | 6 mL |
| Large RNA Removal Column | 25 |
| microRNA Enrichment Column | 25 |
| Collection Tube | 50 |
| Elution Tubes (1.7 mL) | 50 |
| Product Insert | 1 |
Everything you need to run a trial PACE® allele-specific PCR Genotyping Reaction on your existing lab equipment. Each PACE Trial Kit includes Test DNA samples, PACE Genotyping Assays, PACE Master Mix and a comprehensive PACE Genotyping Trial Kit Manual.
Anyone who wants to try PACE genotyping reagents in their lab for the first time with a set of validated DNA samples, SNP assays and PACE Master Mix.
Step 1. Dispense each of the three trial DNA samples (DNA 1, 2 and 3) plus water (No Template Control) in triplicate onto a PCR plate using the suggested volumes.
Step 2. Combine appropriate volumes of PACE Genotyping Master Mix with PACE Genotyping Assay in a tube, as directed, then mix.
Step 3. Dispense the combined mixtures into each of the wells containing DNA using volumes indicated. Each test now contains a complete PACE Genotyping Reaction.
Step 4. Seal your PCR plate with an optically clear seal and centrifuge to ensure all components are at the bottom of the wells.
Step 5.Thermally cycle the reaction plate using the thermal cycling conditions provided.
Step 6. Read the plate and compare data produced with the expected results provided in the manual. Simple!
More information on the PACE genotyping chemistry and how it works can be found here: www.3crbio.com/#pace. PACE allele-specific PCR is used for the detection of SNPs, Indels and other sequence variants.
