Introduction
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 Mini Kit enables efficient purification of these circulating nucleic acids from human plasma, serum, or urine. Samples can be fresh or frozen (provided that they have not been frozen and thawed more than once).
Details
Specifications
| Features | Specifications |
| Main Functions | Isolation circulating DNA from 1ml plasma, serum, body fluids |
| 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 or vacuum) |
| Sample type | Serum, plasma and other cell-free fluid samples |
| Sample amount | 1ml |
| Elution volume | ≥30μl |
| Time per run | ≤50 minutes |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 100µg |
Principle
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).
Advantages
- High yield - most optimal process, free DNA (>50bp) can be obtained to the maximum extent
- High concentration - low elution volume, ensuring high nucleic acid concentration
- High purity - low alcohol binding method, completely removing inhibitor and protein pollution
- High recovery - DNA can be recovered at the level of PG by silica gel column purification
Kit Contents
| Contents | D318102 | D318103 |
| Purification Times | 50 Preps | 250 Preps |
| Buffer ACL | 50 ml | 250 ml |
| Buffer ACB* |
60 ml | 300 ml |
| Buffer DCW1* | 22 ml | 88 ml |
| Buffer DCW2* | 10 ml | 50 ml |
| Proteinase K | 120 mg | 540 mg |
| Protease Dissolve Buffer | 10 ml | 30 ml |
| Carrier RNA | 110 μg | 310 μg |
| Nuclease Free Water |
10 ml | 30 ml |
| HiPure CFDNA Mini Columns |
50 | 250 |
| 2 ml Collection Tubes |
100 | 500 |
Storage and Stability
Proteinase K and 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.
