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 plasma usually 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, body fluids using vacuum protocol |
| Applications | qPCR, liquid or solid chip analysis, hybridization and SNP detection, etc. |
| Purification method | Mini spin column |
| Purification technology | Silica technology |
| Process method | Manual (vacuum) |
| Sample type | Serum, plasma and other cell-free fluid samples |
| Sample amount | 1-5ml |
| Elution volume | ≥50μl |
| Time per run | ≤60 minutes |
| Liquid carrying volume per column | 4ml |
| Binding yield of column | 1mg |
This product is based on silica Column purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption 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.
| Contents | IVD3182 |
| Purification Times | 50 |
| 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 Tube | 50 |
| Vac-Connector | 50 |
Storage and Stability
Proteinase K, Carrier RNA should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 weeks) 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 be stored 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.
Experiment Data
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 plasma usually as short fragments,
DBCO-PEG5-acid is an analog of DBCO-Acid with PEG linker and a DBCO group. DBCO group is a highly reactive cycloalkyne which is commonly used for copper-free Click Chemistry reactions. The hydrophilic PEG chain allows for increased water solubility of compounds in aqueous media. The terminal carboxylic acid can react with primary amine groups in the presence of activators (e.g. EDC, or HATU) to form a stable amide bond. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
DBCO-PEG5-acid is an analog of DBCO-Acid with PEG linker and a DBCO group. DBCO group is a highly reactive cycloalkyne which is commonly used for copper-free Click Chemistry reactions. The hydrophilic PEG chain allows for increased water solubility of compounds in aqueous media. The terminal carboxylic acid can react with primary amine groups in the presence of activators (e.g. EDC, or HATU) to form a stable amide bond. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Both high molecular weight DNA (greater than 1 kb in size; mostly cell associated) and the smaller DNA (150 – 250 bp; derived from the circulation) is effectively isolated and purified using a rapid and convenient spin column protocol. This kit can be used to isolate DNA from a broad range of viruses in urine as well. Salts, metabolic wastes, proteins and other contaminants are removed to yield inhibitor-free DNA for use in sensitive applications. The DNA is of excellent quality for various downstream applications such as PCR, qPCR and DNA fingerprinting, methylation studies and more.
This kit is fully compatible with Norgen’s Urine Collection and Preservation Tubes.
Urine DNA Isolation Kit
This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 50 μL to 1.75 mL of urine. Multiple samples can be processed in 30 minutes.
Urine DNA Isolation Kit (Slurry Format)
This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 3 mL to 25 mL. Multiple samples can be processed in 30 minutes. Multiple samples can be processed in 45 minutes.
Urine DNA Isolation Maxi Kit (Slurry Format)
This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 25 mL of urine up to 80 mL. Multiple samples can be processed in 45 minutes.
Background
DNA found in urine can be divided into 2 basic categories. The larger species, genomic-DNA (gDNA), is generally greater than 1 kb in size, and appears to be derived mainly from exfoliated cells. The second species is smaller, generally between 150 and 250 bp (apoptotic-DNA), and derives, at least in part, from the circulation. The second species is also considered as an RNA/DNA hybrid as reported by Halicka et al. (2000). Both types of DNA can be isolated reliably using this kit.
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| Kit Specifications | |
| Volume of Urine Processed | 1.75 mL |
| Average Yield* | Up to 50 ng |
| Size of DNA Purified | Large (> 1 kb) and small (150-250 bp) |
| Time to Complete 10 Purifications | 30 minutes hands-on time (plus a 1 hour incubation) |
* Yield will vary depending on the type of sample processed
Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. The kit contains a ready-to-use Proteinase K, which is dissolved in a specially prepared storage buffer. The buffered Proteinase K is stable for up to 2 years after the date of shipment when stored at room temperature.
| Component | Cat. 18100 (50 preps) | Cat. 48800 (50 preps) | Cat. 50100 (50 preps) |
|---|---|---|---|
| Binding Solution K | 15 mL | – | – |
| Slurry B1 | – | 18 mL | 18 mL |
| Proteinase K in Storage Buffer | 2 mL | – | – |
| Pronase K in Storage Buffer | 2 mL | – | – |
| Soluton WN | 9 mL | – | – |
| Binding Buffer A | – | – | 50 mL |
| Lysis Buffer A | – | 30 mL | 30 mL |
| Wash Solution A | – | 38 mL | 38 mL |
| Wash Solution B | 30 mL | – | – |
| Wash Solution D | 9 mL | – | – |
| Binding Solution K | 15 mL | – | – |
| Elution Buffer B | 15 mL | 15 mL | 15 mL |
| Micro Spin Columns | 50 | – | – |
| Mini Filter Spin Columns | – | 50 | 50 |
| Collection Tubes | 50 | 50 | 50 |
| Elution Tubes (1.7 mL) | 100 | 100 | 100 |
| Product Insert | 1 | 1 | 1 |