The EXTRACTME DNA BLOOD kit is designed for the rapid and efficient purification of high quality DNA (genomic, mitochondrial and/or viral) from whole blood (fresh or frozen, human or mammalian), plasma, serum, buffy coat, lymphocytes and body fluids.The isolation protocol and buffer formulations were optimized for high isolation efficiency and DNA purity.
up to 1 ml of fresh or frozen blood; up to 200 μl plasma, serum, buffy coat, lymphocytes or body fluids
3-10 μg DNA from 200 μl of human blood
approx. 27 μg DNA
25 minutes (including incubation time)
A260/A280 ratio = 1.7 – 1.9
The DNA purification procedure consists of five steps and utilizes spin minicolumns with membranes, which efficiently and selectively bind nucleic acids. In the first isolation step, the red blood cells are lysed. The cells contain no DNA and are a potential source of PCR inhibitors. They must therefore be separated from the white blood cells prior to DNA isolation. Then the white blood cells are subjected to enzymatic lysis by Proteinase K in optimized BL Buffer. At this stage, all cell membranes and proteins are degraded. After the addition of chaotropic salts, the lysate is applied to the purification minicolumn membrane and the DNA is bound. The two-step washing stage effectively removes impurities and enzyme inhibitors. The purified DNA is eluted using either a low ionic strength buffer (Elution Buffer) or water (pH 7.0-9.0) and may be used directly in all downstream applications such as PCR, qPCR, Southern blotting, DNA sequencing, enzymatic restriction, ligation and so forth, or stored until ready to use.
The quality of each production batch (LOT) of the EXTRACTME DNA BLOOD kit is tested using standard QC procedures. The purified DNA concentration, quality and stability are evaluated by gel electrophoresis and spectrophotometer. In addition, the functional quality is tested by qPCR and digestion with restriction enzymes.
Manufacturer: BLIRT S.A.