The EXTRACTME DNA YEAST kit is designed for the rapid and efficient purification of high quality DNA from broth and plate yeast cultures as well as frozen cells. The isolation protocol and buffer formulations were optimized for high isolation efficiency and DNA purity.
broth or plate yeast culture, frozen cell pellet
- up to 1 x 108 cells → 100%
- 2 x 108 ÷ 9 x 108 cells → 30-70%
approx. 40 μg DNA
approx. 75 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 cell walls of the yeast cells are disintegrated in the YS Buffer (Spheroplast Buffer). Thus obtained spheroplasts are enzymatically lysed by the Proteinase K in the lysis buffer. To obtain an RNA-free DNA sample, the RNA is removed by RNase A. After the addition of chaotropic salts, the lysate is applied to the purification column membrane and the DNA is bound. The two-step washing stage effectively removes impurities and enzyme inhibitors. The purified DNA is eluted using 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 EXTRACTME DNA YEAST 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.