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This kit provides a simple and fast solution for the extraction of circulating nuclear acid from serum, plasma, and other cell-free liquid samples. Circulating nucleic acid refers to the free extracellular nucleic acid produced by cell apoptosis, of which fragments are generally below 1KB. The kit is based on silica gel column purification technology, which is no need for toxic phenol chloroform extraction and time-consuming alcohol precipitation during the extraction. The obtained Circulating Nucleic Acid can be directly used for quantitative PCR, liquid or solid phase chip analysis, hybridization, and SNP detection.
HiPure Circulating DNA/RNA Kit adopts a unique solution system and multiple layers of filter membranes with different pore sizes, which can efficiently process large volumes of serum and plasma samples and capture extremely small amounts of free nucleic acids.
Specifications
| Features | Specifications |
| Main Functions | Isolation both Circulating DNA/RNA (include miRNA) from 1-5 ml serum and plasma |
| Applications | qPCR / RT-PCR, liquid or solid-phasechip analysis, hybridization and SNP detection |
| Purification method | Midi spin column |
| Purification technology | Silica technology, DNA filtration technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | serum, plasma, and other cell-free liquid samples |
| Sample amount | 1-5 ml |
| Elution volume | ≥20μl |
| Time per run | ≤100 minutes |
| Liquid carrying volume per column | 4 ml |
| Binding yield of column | 1 mg |
This kit is based on silica gel column technology. Serum or other liquid samples are lysed and digested in buffer CFL. After adding buffer CFP, the protein is removed by centrifugation to obtain the supernatant. Isopropanol is added to precipitate the total nucleic acid and transferred to the column for filtration. DNA / RNA is adsorbed on the membrane of the column, while the protein is not adsorbed and removed with the filtrate.The column is washed with buffer MGW1 to remove protein and other impurities, and then washed with buffer RW2 to remove salt. Finally, DNA / RNA is eluted by low salt buffer. The eluted DNA / RNA can be directly used for quantitative PCR/ RT-PCR, liquid or solid-phase chip analysis, hybridization and SNP detection.
Advantages
Kit Contents
| Contents | R431602 | D431603 |
| Purification Times | 50 Preps | 250 Preps |
| HiPure RNA Micro Columns | 50 | 5 x 50 |
| HiPure Viral Midi Columns | 50 | 5 x 50 |
| 15 ml Collection Tubes | 50 | 5 x 50 |
| 2ml Collection Tubes | 50 | 5 x 50 |
| Buffer CFL | 150 ml | 2 x 375 ml |
| Buffer CFP | 30 ml | 150 ml |
| Buffer MGW1* | 100 ml | 2 x 250 ml |
| Buffer RW2* | 2 x 50 ml | 5 x 100 ml |
| RNase Free Water | 10 ml | 50 ml |
Storage and Stability
The kit components can be stored at room temperature (15–25°C) and are stable for at least 18 months under these conditions.
This kit provides a simple and fast solution for the extraction of circulating nuclear acid from serum, plasma, and other cell-free liquid samples. Circulating nucleic acid refers to the free extracellular nucleic acid produced by cell apoptosis, of which fragments are generally below 1KB. The kit is based on silica gel column purification technology, which is no need for toxic phenol chloroform extraction and time-consuming alcohol precipitation during the extraction. The obtained Circulating Nucleic Acid can be directly used for quantitative PCR, liquid or solid phase chip analysis, hybridization, and SNP detection.
HiPure Circulating DNA/RNA Kit adopts a unique solution system and multiple layers of filter membranes with different pore sizes, which can efficiently process large volumes of serum and plasma samples and capture extremely small amounts of free nucleic acids.
Norgen’s EXTRAClean Urine Cell-Free Circulating RNA Purification Kits provide a fast, reliable, reproducible, and simple procedure for isolating circulating RNA and exosomal RNA from various urine inputs ranging from 250 μL and up to 30 mL. The purified RNA is of the highest integrity, and can be used in a number of downstream applications including real-time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extensions, and expression array assays.
EXTRAClean Cell-Free Circulating RNA Purification Mini Kit:
For sample volumes ranging from 250 μL to 2 mL
EXTRAClean Cell-Free Circulating RNA Purification Midi Kit:
For sample volumes ranging from 2mL to 10mL. The first column will handle the large volume input of urine that is followed by a concentration on a mini column for a final elution of 50 μL to 100 μL
EXTRAClean Cell-Free Circulating RNA Purification Maxi Kit:
For sample volumes ranging from 10 mL to 30 mL. The first column will handle the large volume input of urine that is followed by a concentration on a mini column for a final elution of 50 μL to 100 μL.
Figure 1 / 3
Click for expanded view
| Kit Specifications | |
|---|---|
| Sample Type | Fresh or frozen urine |
| Sample Volume Range | 10 to 30 mL |
| Minimum Elution Volume | 50 μL |
| Maximum Elution Volume | 100 μL |
| Time to Complete 10 Purifications | 40–50 minutes |
| Size of RNA Purified | All sizes, including miRNA and small RNA (<200 nt) |
| Average Yields ¥ | Variable depending on specimen |
¥Please check page 5 of the protocol for average urine yields and common RNA quantification methods.
All buffers should be kept tightly sealed and stored at room temperature. These kits are 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 | Mini (Cat. 72900) | Midi (Cat. 72910) | Maxi (Cat. 72920) |
|---|---|---|---|
| Binding Solution K | 25 mL | 75 mL | 1 x 75 mL 1 x 25 mL |
| Lysis Buffer A | 30 mL | 20 mL | 20 mL |
| Wash Solution A | 18 mL | 18 mL | 18 mL |
| Elution Solution A | 6 mL | 6 mL | 6 mL |
| Collection Tubes | 50 | 20 | 10 |
| Elution Tubes (1.7 mL) | 50 | 20 | 10 |
| EXTRAClean Mini Spin Columns | 50 | 20 | 10 |
| EXTRAClean Midi Spin Columns | – | 20 | – |
| EXTRAClean Maxi Spin Columns | – | – | 10 |
| Product Insert | 1 | 1 | 1 |
Propargyl-PEG6-amine is a crosslinker with a hydrophilic PEG spacer which can increase the hydrophilicity of the molecule in aqueous environment. The amine group is reactive with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde) etc. The propargyl group can react with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Propargyl-PEG6-amine is a crosslinker with a hydrophilic PEG spacer which can increase the hydrophilicity of the molecule in aqueous environment. The amine group is reactive with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde) etc. The propargyl group can react with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
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