Extracta Plus RNA
- Rapid Protocol – From cells or tissue samples to high-quality total RNA in just 25 mins
- Streamlined gDNA removal – Novel DNA removal column eliminates gDNA contamination and need for DNase treatment
- Extraction of low input quantities – Ability to extract RNA from as little as 10 cells
- Downstream compatibility – Ideal for applications such as RT-PCR, RT-qPCR and next generation sequencing
Extracta Plus RNA Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention or treatment of a disease.
Description
The Extracta Plus RNA kit provides rapid extraction and purification of RNA from cultured human or animal cells and from various tissue samples. The Extracta Plus spin column workflow enables simultaneous processing of multiple samples in 25 mins to yield purified RNA suitable for a range of downstream applications. The kit includes specially designed Extracta Plus DNA removal columns for effective removal of genomic DNA contamination. This provides a convenient solution for applications that are sensitive to low amounts of DNA contamination such as RT-qPCR and NGS.
Details
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Volume |
|
|
Component Description |
10 Reactions |
50 Reactions |
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Buffer EPRL |
6 ml |
45 ml |
|
Buffer EPRW1 |
7 ml |
45 ml |
|
Buffer EPRW2 |
3 ml |
11 ml |
|
RNase-free water |
10 ml |
10 ml |
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Extracta Plus Spin Column |
10 piece |
50 piece |
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Extracta Plus DNA Removal Column |
10 piece |
50 piece |
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Collection Tubes 1.5 ml |
10 piece |
50 piece |
|
Collection Tubes 2 ml |
10 piece |
50 piece |
Performance Data
Extracta Plus RNA complete workflow
This procedure can be completed in just 25 minutes, resulting in ready-to-use total RNA applicable for sensitive downstream applications such as RT-qPCR and NGS.
Total RNA purification yield
Quantitative extraction of RNA from dilute cell quantities. Total RNA was purified in triplicate from HeLa cell quantities from as much as 100,000 to as little as 10 cells using the Extracta Plus RNA Kit and eluted in a volume of 30 µl. A portion of the RNA (1.5 µl) was then used for qPCR analysis of beta-actin mRNA using the qScript XLT ToughMix. Results demonstrate successful and reproducible extraction of RNA from a wide range of input.
Effective RNA yield with negligible gDNA
Total RNA was purified from (A) 1 x 106HeLa cells or (B) 10 mg rat kidney tissue using the Extracta RNA Plus Kit. RT-qPCR assays were performed with (+RT) or without (-RT) reverse transcriptase.
Long mRNA target amplification
Extraction of intact high molecular weight RNA from tissues and cultured cells. Total RNA was purified from either 5 or 10 mg of the rat kidney (K) or lung (L) tissue or from 1X106 cultured HeLa cells (H) using the Extracta RNA Plus Kit. Large fragments of the rat Dynein mRNA and the human SYNE1 mRNA were amplified via RT-PCR from 200 ng of RNA using qScript Ultra Flex Kit and then repliQa HiFi Toughmix. Gel analysis of the product demonstrates large quantities of intact and high molecular weight mRNA originally present in the RNA samples.
Typical yields from common sources
Typical yields from common sources.
Effective gDNA removal
qPCR assays for human or rat β-actin were performed. A standard curve of human/rat gDNA was used to quantify the amount of gDNA contamination in the purified RNA. For all samples, gDNA was present in negligible quantities.