This kit is for the extraction of high purity total RNA from biological samples such as cultured cells and tissues.
The recovered total RNA can be used mainly in enzymatic reactions such as RT-PCR.
As this kit employs the principle that magnetic silica beads bind total RNA present in a lysate solution, it is not necessary to perform deproteinization using harmful reagents such as phenol, ethanol precipitation, or high-speed centrifugation, which are necessary using the conventional AGPC method, thus simplifying the extraction process.
This kit is suitable for the MFX Series automatic nucleic acids extraction system and can also be used as a manual kit for B/F (solid-liquid) separation using a magnetic beads separation stand.
1. Examination of Yield and Purity
Total RNA was extracted from 2 x 106 HeLa cells using this kit and the widely used AGPC RNA extraction method.
The yield obtained with this kit was approximately equivalent to that obtained by AGPC method.
With regard to purity, the electrophoretic patterns and OD ratios indicate that this kit extracts high purity RNA with almost no contamination by impurities such as proteins and DNA.
2. Example PCR with Extracted RNA
1 g of the total RNA extracted from 2 x 106 HeLa cells was used to perform RT-PCR with transferrin receptor mRNA as a target (2115 bp).
As a result, amplification of the target was detected only in the reverse transcription reactions, and expression of the normally low-volume expression transferrin receptor gene was confirmed.
The result showed that using RNA obtained with this kit enables RT-PCR with various targets.
3. Extraction from Rat Tissue
Total RNA was extracted from rat tissue using this kit.
50-100 mg of rat tissue was frozen in liquid nitrogen and crushed with a hammer. 750-900 µl of ice-cold Lysis & Binding Solution (2-ME content) was added, and the mixture homogenized completely with a pestle. The supernatant was removed and left at room temperature for 10 minutes to extract the total RNA. The results showed that by performing homogenization, using this kit enabled the extraction of high quality RNA, even from tissue.
This kit extracts high purity total RNA according to the following steps.
MagExtractor -Viral RNA-
To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.