This kit provides for rapid spin column isolation and purification of total RNA and genomic DNA simultaneously from a single plant sample without splitting the lysate. Norgen’s plant lysis solution is highly robust and effective over a wide range of plants including challenging samples. The total RNA and genomic DNA are both column purified in under 30 minutes. Since RNA and DNA are isolated without splitting the lysate, variability and inconsistent results are reduced. All sizes of RNA including microRNA are recovered without the need for phenol. Optional on-column DNase and RNase treatments provide flexibility to isolate DNA-free RNA or RNA-free DNA respectively. Isolated nucleic acids are of a high quality and yield, and are ready for downstream use including PCR, qPCR, RT-PCR, qRT-PCR, sequencing and more.
Background
It is often necessary to isolate total RNA and genomic DNA from a single plant sample, such as for studies of gene expression, mutant or transgenic plant characterization, and host plant-pathogen characterization. This is of great benefit when isolating RNA and DNA from precious, difficult to obtain or very small samples. Furthermore, gene expression analysis will be more reliable since the RNA and DNA are derived from the same sample, therefore eliminating inconsistent results.
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| Kit Specifications | |
| Column Binding Capacity | 50 μg for RNA 15 μg for genomic DNA |
| Maximum Column Loading Volume | 650 μL |
| Size of RNA Purified | All sizes, including small RNA (< 200 nt) |
| Maximum Amount of Starting Material: Plant Tissues Plant Cells | 100 mg 5 x 106 |
| Time to Complete 10 Purifications | 30 minutes |
| Average Yields* Peach Leaves (100 mg) | 40 μg RNA, 5 μg gDNA |
* Yield will vary depending on the type of sample processed
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 2 years in their unopened containers.
| Component | Cat. 24400 (50 preps) |
|---|---|
| Lysis Buffer M | 40 mL |
| Binding Buffer I | 7 mL |
| Solution WN | 18 mL |
| Wash Solution A | 38 mL |
| Elution Buffer E | 20 mL |
| Enzyme Incubation Buffer B | 6 mL |
| Filter Columns | 50 |
| Spin Columns | 50 |
| Collection Tubes | 100 |
| Elution Tubes (1.7 mL) | 50 |
| Product Insert | 1 |
These kits provide a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating and exosomal RNA using a convenient spin column method. These kits can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used, as heparin can significantly interfere with many downstream applications such as RT-PCR. The purified plasma/serum RNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, and NGS. The EXTRAClean columns undergo stringent processing and rigorous quality control measures to minimize contamination traces, ensuring optimal results for sensitive applications such as NGS.
Cell-free circulating RNA, including exosomal RNA in plasma or serum, has the potential to provide biomarkers for certain cancers and disease states, and includes tumor-specific extracellular RNA in the blood. Exosomes are 40 – 100 nm membrane vesicles, which are secreted by most cell types. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascitic fluids, among other biological fluids. Evidence has been accumulating recently that these vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. The exosomes contain cell-specific proteins, lipids and RNAs, which are transported to other cells, where they can alter function and/or physiology. These exosomes may play a functional role in mediating adaptive immune responses to infectious agents and tumours, tissue repair, neural communication and transfer of pathogenic proteins. Recent work has demonstrated the presence of distinct subsets of microRNAs within exosomes which depend upon the tumour cell type from which they are secreted. For this reason, exosomal RNAs may serve as biomarkers for various diseases including cancer. As the RNA molecules encapsulated within exosomes are protected from degradation by RNAses, they can be efficiently recovered from biological fluids, such as plasma or serum.
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 50 μL to 200 μL. The purified plasma/serum RNA is eluted in a flexible final volume of 10 μL to 25 μL.
This utilizes a two-column method, and can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 250 μL to 1.5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 μL to 100 μL.
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 2 mL to 5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 μL to 100 μL.
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| Kit Specifications | |
| Sample Type | Plasma/Serum |
| Anti-Coagulant (for Plasma)† | EDTA or Citrate |
| Sample Volume Range | 50 to 200 μL |
| Minimum Elution Volume | 10 μL |
| Maximum Elution Volume | 25 μL |
| Time to Complete 10 Purifications | 15 – 20 minutes |
| Size of RNA Purified | All sizes, including miRNA and small RNA (<200 nt) |
| Average Yields¥ | Variable depending on specimen |
† This kit is suitable for the isolation of RNA from fresh or frozen serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR
¥ Please check page 5 for Average Plasma/Serum Yields and Common RNA Quantification Methods
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.
| Component | Cat. 73700 (50 preps) | Cat. 73710 (20 preps) | Cat. 73720 (10 preps) |
|---|---|---|---|
| Lysis Buffer A | 2 x 20 mL | 100 mL | 1 x 130 mL 1 x 30 mL |
| Wash Solution A | 18 mL* | 1 x 38 mL* 1 x 18 mL* | 38 mL* |
| Elution Solution A | 6 mL | 6 mL | 6 mL |
| Elution Buffer F | – | – | 15 mL |
| EXTRACLean Micro Spin Columns | 50 | – | – |
| EXTRACLean Mini Spin Columns | – | 20 | 10 |
| EXTRACLean Midi Spin Columns | – | 20 | – |
| EXTRACLean Maxi Spin Columns | – | – | 10 |
| Collection Tubes | – | 20 | 10 |
| Elution Tubes (1.7 mL) | 50 | 20 | 10 |
| Product Insert | 1 | 1 | 1 |
This kit is designed for the rapid preparation of genomic DNA from various tissue samples, cultured cells, viruses, bodily fluids and swabs using a rapid spin column protocol. Purified DNA is of an excellent yield and quality, and is immediately ready for any downstream application including PCR, qPCR, genotyping, sequencing and more. The protocol can be completed in approximately 80 minutes (including incubation time).
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| Kit Specifications | |
| Column Binding Capacity | 25 µg |
| Average Yield:* HeLa Cells (1 x 106) Tissue (from 10 mg kidney) | 8 µg 10 µg |
| Maximum Amount of Starting Material: Animal Tissues Cultured Cells Bodily Fluids (blood, saliva) Viral Suspension | 20 mg 3 x 106 cells 150 µL 150 µL |
| Time to Complete 10 Purifications | 80 minutes |
* Yield will vary depending on the type of sample processed
Storage Conditions and Product Stability
The Proteinase K should be stored at -20°C upon arrival and after reconstitution. All other solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.
| Component | Cat. 24700 (50 preps) | Cat. 24750 (100 preps) | Cat. 24770 (250 preps) |
|---|---|---|---|
| Digestion Buffer A | 25 mL | 2 x 25 mL | 5 x 25 mL |
| Buffer SK | 30 mL | 2 x 30 mL | 5 x 30 mL |
| Wash Solution A | 18 mL | 2 x 18 mL | 5 x 18 mL |
| Elution Buffer B | 30 mL | 2 x 30 mL | 5 x 30 mL |
| Proteinase K | 12 mg | 2 x 12 mg | 5 x 12 mg |
| Spin Columns | 50 | 100 | 250 |
| Collection Tubes | 50 | 100 | 250 |
| Elution Tubes (1.7 mL) | 50 | 100 | 250 |
| Product Insert | 1 | 1 | 1 |