This kit is for the extraction of high purity genomic DNA from biological samples such as blood and cultured cells. The recovered genomic DNA can be used directly, mainly in enzymatic reactions such as PCR.
As this kit employs the principle that magnetic silica beads bind genomic DNA present in a lysate solution, it is not necessary to perform deproteinization using harmful reagents such as phenol, ethanol precipitation, or high-speed centrifugation, 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.
When extracting genomic DNA from blood, whole blood can be used for DNA extraction, doing away with the need for leukocyte separation as required by conventional methods.
1. Examination of Yield and Purity
Genomic DNA was extracted from 100 µl of whole blood using this kit and others commercially available. The yield obtained with this kit was approximately equivalent to those obtained with the commercially available ones. With regard to purity, impurities such as proteins or RNA were more likely to have been present with use of the other commercially available kits (companies A and C), whereas such impurities were not detected with use of this kit. The results show that use of this kit allows extraction of high purity genomic DNA from biological samples such as whole blood.
2. Example PCR Using Extracted DNA
After extracting genomic DNA from 100 l of whole blood collected from ten healthy, unrelated donors using this kit, we carried out PCR. Using 1/20 of the total amount of the extracted DNA, PCR was performed with Taq DNA polymerase with a single-locus probe of the VNTR regions of DNA, MCT 118, as a target. Approximately 1.4-2.2 µg of high purity genomic DNA with an A260/280 ratio between 1.83 and 1.84 was obtained.
MCT118 is a repeated sequence of 16 bases, and it is thought that by determining the number of these repeats profiling can be performed. In this PCR with MCT118 as a target, the specific bands for each individual were detected.
This kit extracts high purity genomic DNA according to the following steps.
1. Add Lysis & Binding Solution containing chaotropic reagents to the sample to dissolve the cells.
2. Add Magnetic Silica Beads and mix for the DNA in the sample to adhere to the silica’s surface.
3. Wash the beads with the two kinds of Washing Solution and remove unwanted proteins, RNA, salts, etc.
4. Elute and recover DNA from the beads with sterile water or low salt buffer, etc.
MagExtractor -Plant Genome-
To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.