Introduction
Free-circulating nucleic acids, such as tumor-specific extracellular nucleic acid fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasma usually as short fragments, <1000bp(Nucleic Acid). The HiPure Circulating Nucleic acid Micro Kit enables efficient purification of these circulating nucleic acids from human plasma, serum, or urine. Samples can be either fresh or frozen (provided that they have not been frozen and thawed more than once). Free-circulating cell-free DNA, RNA or viral nucleic acids are eluted in Nuclease Free Water, ready for use in amplification reactions or storage at -30 to -15°C. Purified nucleic acids are free of proteins, nucleases, and other impurities.
Details
Specifications
| Features | Specifications |
| Main Functions | Isolation circulating DNA from 0.6ml 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 | 0.6ml |
| Elution volume | ≥30μl |
| Time per run | ≤40 minutes |
| Liquid carrying volume per column | 800μl |
| Binding yield of column | 100μg |
Principle
This product is based onsilica column purification. The sample is lysed and digested with lysate andprotease, DNA is released into the lysate. Transfer to an adsorption plate andfilter column. Nucleic acid is adsorbed on the membrane, while protein is notadsorbed and is removed with filtration. After washing proteins and otherimpurities, Nucleic acid was finally eluted with low-salt buffer.
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 recoveredat the level of PG by silica gel column purification
Kit Contents
| Contents | D318002 | D318003 |
| Purification Times | 50 Preps | 250 Preps |
| Buffer ACL | 40 ml | 200 ml |
| Buffer DCW1 | 22 ml | 110 ml |
| Buffer DCW2* | 20 ml | 2 x 50 ml |
| Proteinase K | 34 mg | 180 mg |
| Protease Dissolve Buffer | 1.8 ml | 10 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 | 5 x 100 |
Storage and Stability
Proteinase K, Carrier RNAshould 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. Theremaining kit components can be stored dry at room temperature (15-25°C) andare 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.
