30,00 € – 503,00 €The EXTRACTME DNA YEAST KIT is designed for a 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. The product is intended for research use only.
DNA purification procedure consists of five steps and utilizes spin minicolumns with membranes which efficiently and selectively bind nucleic acids. During the first step cell walls of the yeast cells are disintegrated in the YS Buffer (Spheroplast Buffer). Spheroplasts obtained this way are enzymatically lysed by Proteinase K in lysis buffer. To obtain an RNA-free DNA sample, RNA is removed by RNase A. After the addition of chaotropic salts, the lysate is applied to the purification column membrane and DNA is bound. The two-step washing stage effectively removes impurities and enzyme inhibitors. Purified DNA is eluted with then use of 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.
Broth or plate yeast culture, frozen cell pellet
Up to 1 x 108 cells → 100%
2 x 108 ÷ 9 x 108 → 30–70%
Approx. 40 μg DNA
Approx. 75 minutes (including incubation time)
A260/A280 ratio = 1.7 – 1.9
RECOMMENDATIONS AND IMPORTANT NOTES
DNA isolation efficiency and purity can depend not only on the number of yeast cells, cell type (morphology) or antibiotics in the grow medium, but also on the age and condition of the yeast cells.
The kit is designed to isolate DNA from max. 1 x 108 yeast cells. Using greater number of cells may significantly reduce the isolation efficiency and the purity of the DNA.
Extracting DNA from fresh starting material or frozen cell pellets is recommended. To minimize DNA degradation, avoid subjecting the sample material to repeated freeze/thaw cycles. Using old or repeatedly frozen/thawed material may result in low efficiency isolation of high molecular DNA.
Protocol & Manual
EXTRACTME DNA YEAST KIT (EM10) - Manual
EXTRACTME DNA YEAST KIT – TROUBLESHOOTING
Quality & Safety
The quality of each production batch (LOT) of the EXTRACTME DNA YEAST KIT is tested with the use of standard QC procedures. Purified DNA concentration and quality are evaluated by gel electrophoresis and spectrophotometer.
EXTRACTME DNA YEAST KIT (EM10) - MSDS
The influence of different pH on the electrophoretic behaviour of Saccharomyces cerevisiae modified by calcium ions
Authors: Agnieszka Rogowska, Paweł Pomastowski, Michał Złoch, Viorica Railean-Plugaru, Anna Król, Katarzyna Rafińska, Małgorzata Szultka-Młyńska & Bogusław Buszewski Products: EXTRACTME DNA YEAST KIT (EM10) Year: 2018 Source: Scientific Reportsvolume 8, Article number: 7261 (2018)
A new assay based on terminal restriction fragment length polymorphism of homocitrate synthase gene fragments for Candida species identification
Authors: Kasjan Szemiako, Anna Śledzińska, Beata Krawczyk Products: EXTRACTME DNA YEAST KIT (EM10) Year: 2017 Source: J Appl Genetics (2017) 58:409–414
Isolation and Characterization of Phenol-Degrading Psychrotolerant Yeasts
Authors: Natalia Filipowicz, Malwina Momotko, Grzegorz Boczkaj, Tomasz Pawlikowski, Marta Wanarska, Hubert Cieśliński Products: EXTRACTME DNA YEAST KIT (EM10) Year: 2017 Source: Water Air Soil Pollut (2017) 228: 210