These kits provide a fast, reliable and convenient spin column method for the isolation of high quality, high purity and inhibitor-free cell-free circulating DNA (cfc-DNA) from plasma/serum sample. These kits are designed to isolate all sizes of cfc-DNA from either fresh or frozen plasma/serum samples and the purified DNA is eluted into a flexible elution volume ranging from 25 µL to 50 µL. The purified plasma/serum cfc-DNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive PCR and Southern Blot analysis, microarrays and NGS.
Background
Plasma/Serum cell-free circulating DNA (cfc-DNA) has the potential to provide biomarkers for certain cancers and disease states as well as fetal DNA in maternal blood. Currently, significant advancements are being made in utilizing cfc-DNA as biomarkers for the early diagnosis, prognosis and monitoring of therapy for several cancer types and autoimmune diseases. Cell-free mitocondrial DNA (cfmtDNA) is also under investigation for its clinical significance. This cfc-DNA is usually present as short fragments of less than 1000 bp. In addition, cell-free fetal DNA has been widely used as a non-invasive method for prenatal diagnosis including early identification of fetal sex, genetic studies for families at high risk for inherited genetic disorders, screening for Rhesus factor, screening for aneuploidy and identification of preeclampsia.
Plasma/Serum Cell-Free Circulating DNA Purification Micro Kit Dx
For serum input volumes 10 µL – 200 µL. Purify high-quality DNA in 15-20 minutes
Plasma/Serum Cell-Free Circulating DNA Purification Mini Kit Dx
For serum input volumes 200 µL – 500 µL. Purify high-quality DNA in 15-20 minutes.
Plasma/Serum Cell-Free Circulating DNA Purification Midi Kit Dx
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 25 µL to 50 µL. All components for the purification & concentration are provided in one convenient & fast kit for the easy processing of large input volumes of bodily fluids. For a schematic workflow of the protocol click here to view. For serum input volumes 1 mL – 4 mL. Purify high-quality DNA in 90 minutes.
Plasma/Serum Cell-Free Circulating DNA Purification Maxi Kit Dx
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 25 µL to 50 µL. All components for the purification & concentration are provided in one convenient & fast kit for the easy processing of large input volumes of bodily fluids. For a schematic workflow of the protocol click here to view. For serum input volumes 5 mL – 10 mL. Two kits in one – purify and concentrate DNA from bodily fluids using a convenient two column system.
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| Kit Specifications | |
| Minimum Plasma/Serum Input | 10 μL |
| Maximum Plasma/Serum Input | 200 μL |
| Size of DNA Purified | ≥ 50 bp |
| Elution Volume | 25-50 μL |
| Time to Complete 10 Purifications | 15-20 minutes |
| Average Yields* | Variable depending on specimen |
*Please check page 7 in the product manual for the Plasma/Serum Average Yields and the Common DNA Quantification Methods.
Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature (15-25°C) for up to 2 years without showing any reduction in performance. Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Kits contain ready-to-use Proteinase K solution, which is dissolved in a specially prepared storage buffer. The Proteinase K is stable for up to 2.5 years after delivery when stored at room temperature. To prolong the lifetime of Proteinase K, storage at 2–8°C is recommended.
| Component | Cat. Dx55100 (50 preps) | Cat. Dx55500 (50 preps) | Cat. Dx55600 (20 preps) | Cat. Dx55800 (10 preps) |
|---|---|---|---|---|
| Binding Buffer B | 2 x 40 mL | 40 mL | 2 x 85 mL 1 x 12 mL | 2 x 85 mL 1 x 40 mL |
| Proteinase K | 2 x 1 mL | 0.6 mL | 6.5 mL | 8 mL |
| Solution WN | 18 mL | 18 mL | 18 mL | 18 mL |
| Wash Solution A | 38 mL | 18 mL | 18 mL | 18 mL |
| Elution Buffer B | 8 mL | 8 mL | 30 mL | 30 mL |
| Micro Spin Columns | 50 | 50 | 20 | 10 |
| Mini Spin Columns | 50 | – | – | – |
| Midi Spin Columns | – | – | 20 | – |
| Maxi Spin Columns | – | – | – | 10 |
| Collection Tubes | 100 | 50 | 20 | 10 |
| Elution Tubes (1.7 mL) | 100 | 50 | 20 | 10 |
| Product Insert | 1 | 1 | 1 | 1 |
PEG3-bis(Amino-Tri-(Propargyl-PEG2-ethoxymethyl)-methane) is a crosslinker consisting of six propargyl groups. The propargyl groups can form triazole linkage with azide-bearing compounds or biomolecules via copper catalyzed Click Chemistry. The hydrophilic PEG spacer increases solubility in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
PEG3-bis(Amino-Tri-(Propargyl-PEG2-ethoxymethyl)-methane) is a crosslinker consisting of six propargyl groups. The propargyl groups can form triazole linkage with azide-bearing compounds or biomolecules via copper catalyzed Click Chemistry. The hydrophilic PEG spacer increases solubility in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
This product is suitable for rapid extraction of RNA (include miRNA) from tissue, cells, blood, s and other clinical samples. RNA can be used directly for RT-PCR, quantitative RT-PCR, test of virus DNA and so on.
Specifications
| Features | Specifications |
| Main Functions | Isolation total RNA（miRNA）from tissue, cell using two columns and DNase plus reagent |
| Applications | RT-PCR, cDNA synthesis, second generation sequencing |
| Products | RNA, miRNA |
| Purification method | Mini spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Clinical tissues, cells, lymphocytes |
| Sample amount | Tissue: <20 mgCells: <5 x 106 |
| Yield | 2-50μg |
| Elution volume | ≥30μl |
| Time per run | ≤25 minutes |
This product is based on silica column purification. Remove paraffin by Buffer DPS. Sample lysis withproteinase K digestion requires only 15 minutes. After lysis, samples are incubated at 80ºC for 15 minutes. Transfer to an adsorption column and RNA is adsorbed on the membrane, while protein is not adsorbed andis removed with filtration. After washing proteins and other impurities, RNA was finally eluted with low-saltbuffer.
Advantages
Kit Contents
| Contents | IVD4121 |
| Purification Times | 50 Preps |
| HiPure DNA Mini Column Ⅱ | 50 |
| HiPure RNA Mini Columns | 50 |
| 2ml Collection Tubes | 150 |
| Proteinase K | 24 mg |
| Protease Dissolve Buffer | 1.8 ml |
| DNase I | 600 μl |
| DNase Buffer | 6 ml |
| Buffer RTL | 40 ml |
| RNA Digestion Buffer | 15 ml |
| Buffer RWC* | 20 ml |
| Buffer RW2* | 20 ml |
| Nuclease Free Water | 10 ml |
Storage and Stability
Proteinase K should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, Proteinase K up to 8 weeks) at room temperature (15–25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15–25°C) and are stable for at least 18 months under these conditions.
This product is suitable for rapid extraction of RNA (include miRNA) from tissue, cells, blood, s and other clinical samples. RNA can be used directly for RT-PCR, quantitative RT-PCR, test of virus DNA and so on.