Free-circulating nucleic acids, such as tumor-specific extracellular DNA fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasmausually as short fragments, <1000bp(DNA). HiPure Circulating DNA Midi Kit enables efficient purification of these circulating nucleic acids from human plasma, serum, or urine. The extracted products can be used for clinical in vitro detection.
Specifications
| Features | Specifications |
| Main Functions | Isolation circulating DNA from 1-5ml plasma, serum, bodyfluids by spin |
| Applications | qPCR, liquid or solid chip analysis, hybridization and SNP detection, etc. |
| Purification method | Mini spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation) |
| Sample type | Serum, plasma and other cell-free fluid samples |
| Sample amount | 1-5ml |
| Elution volume | ≥50μl |
| Time per run | ≤70 minutes |
| Liquid carrying volume per column | 4ml |
| Binding yield of column | 1mg |
This product is based on silica gel purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption plate and filter column. Nucleic acid is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10 Mm Tris,pH 8.0).
Kit Contents
| Contents | D318203D |
| Purification Times | 50 Preps |
| Buffer ACL | 250 ml |
| Buffer ACB* | 300 ml |
| Buffer DCW1* | 22 ml |
| Buffer DCW2* | 10 ml |
| Proteinase K | 540 mg |
| Protease Dissolve Buffer | 30 ml |
| Carrier RNA | 110 μg |
| Nuclease Free Water | 20 ml |
| HiPure CFDNA Mini Columns | 50 |
| 2 ml Collection Tubes | 100 |
| Extender Tubes | 50 |
| Sealing O-Ringes | 50 |
| 50ml Collection Tube | 50 |
| Support Tube | 50 |
Storage and Stability
Proteinase K, Carrier RNA should be stored at 2-8°C upon arrival. However, short-term storage (up to 12weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. The entire kit can bestored at 2-8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.
Free-circulating nucleic acids, such as tumor-specific extracellular DNA fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasmausually as short fragments,
This kit provides a rapid, single column method for the isolation and purification of total RNA (including miRNA) and proteins sequentially from a single sample of cultured animal cells, tissues, blood, bacteria, yeast, fungi or plants. The total RNA and proteins are both column purified in under 25 minutes using a single column.
Purified RNA is of a high quality and yield, and is suitable for NGS, RT-qPCR and microarrays.
This kit eliminates DNA efficiently using a gDNA removal column.
Proteins are eluted in buffer and are ready for downstream applications such as Western Blots, Mass Spec and ELISA. The proteins will not require precipitation, resuspending of pellets, or any further cleaning.
This kit is ideal for researchers who are interested in studying the transcriptome and proteome of a single sample, such as for studies of microRNA profiling, gene expression including gene silencing experiments or mRNA knockdowns, studies involving biomarker discovery, and for characterization of cultured cell lines. Norgen’s RNA/Protein Purification Plus Kit is especially useful for researchers who are isolating macromolecules from precious, difficult to obtain or small samples such as biopsy materials or single foci from cell cultures, as it eliminates the need to fractionate the sample. Furthermore, analysis will be more reliable since the RNA and proteins are derived from the same sample, thereby eliminating inconsistent results. The purified macromolecules are of the highest purity and can be used in a number of different downstream applications.
Protocol
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| Kit Specifications | |
| Maximum Column Binding Capacity | 50 μg for RNA |
| Maximum Column Binding Capacity | 200 μg for Protein |
| Maximum Column Loading Volume | 650 μL |
| Size of RNA Purified | All sizes, including small RNA (< 200 nt) |
| Time to Complete 10 Purifications | 30 minutes |
| Average Yields*: HEK 293 Cells (1 x 106 cells) HEK 293 Cells (1 x 106 cells) Liver (15 mg) Liver (15 mg) | 10-15 μg RNA 70-100 μg protein 30-35 μg RNA 100-150 μg protein |
* Average yields will vary depending upon a number of factors including species, growth conditions used and developmental stage.
Storage Conditions and Product Stability
The Protein Loading Dye should be stored at -20°C after the addition of DL-Dithiothreitol (DTT). 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. 48200 (50 preps) |
|---|---|
| Buffer SKP | 40 mL |
| Wash Solution A | 38 mL |
| Elution Solution A | 6 mL |
| Wash Solution C | 30 mL |
| Binding Buffer A | 8 mL |
| Elution Buffer C | 4 mL |
| Protein Neutralizer | 4 mL |
| Protein Loading Dye | 2 mL |
| gDNA Removal Columns | 50 |
| RNA/Protein Purification Columns | 50 |
| Collection Tubes | 150 |
| Elution Tubes (1.7 mL) | 100 |
| Product Insert | 1 |
Propargyl-PEG6-t-butyl ester is a heterobifunctional linker consisting of an alkyne group and a t-butyl protected carboxyl group. The alkyne group can participate in copper catalyzed azide-alkyne Click Chemistry to form a stable triazole linkage. The t-butyl group can be hydrolyzed under acidic conditions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Propargyl-PEG6-t-butyl ester is a heterobifunctional linker consisting of an alkyne group and a t-butyl protected carboxyl group. The alkyne group can participate in copper catalyzed azide-alkyne Click Chemistry to form a stable triazole linkage. The t-butyl group can be hydrolyzed under acidic conditions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.