Introduction
This kit provides a simple and fast solution for the extraction of circulating nuclear acid from serum, plasma, and other cell-free liquid samples. Circulating nucleic acid refers to the free extracellular nucleic acid produced by cell apoptosis, of which fragments are generally below 1KB. The kit is based on silica gel column purification technology, which is no need for toxic phenol chloroform extraction and time-consuming alcohol precipitation during the extraction. The obtained Circulating Nucleic Acid can be directly used for quantitative PCR, liquid or solid phase chip analysis, hybridization, and SNP detection.
HiPure Circulating DNA/RNA Kit adopts a unique solution system and multiple layers of filter membranes with different pore sizes, which can efficiently process large volumes of serum and plasma samples and capture extremely small amounts of free nucleic acids.
Details
Specifications
Features | Specifications |
Main Functions | Isolation both Circulating DNA/RNA (include miRNA) from 1-5 ml serum and plasma |
Applications | qPCR / RT-PCR, liquid or solid-phasechip analysis, hybridization and SNP detection |
Purification method | Midi spin column |
Purification technology | Silica technology, DNA filtration technology |
Process method | Manual (centrifugation or vacuum) |
Sample type | serum, plasma, and other cell-free liquid samples |
Sample amount | 1-5 ml |
Elution volume | ≥20μl |
Time per run | ≤100 minutes |
Liquid carrying volume per column | 4 ml |
Binding yield of column | 1 mg |
Principle
This kit is based on silica gel column technology. Serum or other liquid samples are lysed and digested in buffer CFL. After adding buffer CFP, the protein is removed by centrifugation to obtain the supernatant. Isopropanol is added to precipitate the total nucleic acid and transferred to the column for filtration. DNA / RNA is adsorbed on the membrane of the column, while the protein is not adsorbed and removed with the filtrate.The column is washed with buffer MGW1 to remove protein and other impurities, and then washed with buffer RW2 to remove salt. Finally, DNA / RNA is eluted by low salt buffer. The eluted DNA / RNA can be directly used for quantitative PCR/ RT-PCR, liquid or solid-phase chip analysis, hybridization and SNP detection.
Advantages
- High yield – most optimized process to obtain maximum free RNA and small RNA
- High concentration – low elution volume (>20μl) to ensure nucleic acid concentration
- High purity – low alcohol combination, completely remove inhibitor and protein pollution
- High recovery – silica gel column technology can recover nucleic acid molecules at the level of PG
- Large volume – 1-2ml serum and plasma samples can be processed at one time
Kit Contents
Contents | R431602 | D431603 |
Purification Times | 50 Preps | 250 Preps |
HiPure RNA Micro Columns | 50 | 5 x 50 |
HiPure Viral Midi Columns | 50 | 5 x 50 |
15 ml Collection Tubes | 50 | 5 x 50 |
2ml Collection Tubes | 50 | 5 x 50 |
Buffer CFL | 150 ml | 2 x 375 ml |
Buffer CFP | 30 ml | 150 ml |
Buffer MGW1* | 100 ml | 2 x 250 ml |
Buffer RW2* | 2 x 50 ml | 5 x 100 ml |
RNase Free Water | 10 ml | 50 ml |
Storage and Stability
The kit components can be stored at room temperature (15–25°C) and are stable for at least 18 months under these conditions.