Description :

Introduction

The EZgene^TM Yeast gDNA Kit allows rapid and reliable isolation of high-quality total cellular DNA from a wide variety of yeast species. Up to 3 mL of log-phase culture (OD₆₀₀ of 1.0 in YPD medium) can be processed. The system combines the reversible nucleic acid-binding properties of ezBind matrix with the speed and versatility of spin column technology to yield approximately 15-30 μg of DNA with an A₂₆₀/A₂₈₀ ratio of 1.7-1.9. Purified DNA is suitable for PCR, restriction digestion, and hybridization techniques. There are no organic extractions thus reducing plastic waste and hands-on time to allow multiple samples to be processed in parallel. EZgene^TM Yeast gDNA Kit will isolate all cellular DNA, including plasmid DNA.

Overview

If using the EZgene^TM Yeast gDNA Kit for the first time, please read this manual before beginning the procedure. Yeast cells are grown to log-phase and speroblasts are subsequently prepared. Following lysis, binding conditions are adjusted and the sample applied to an ezBind DNA Mini column. Two rapid wash steps remove trace salt and protein contaminants and finally DNA is eluted in water or low ionic strength buffer. Purified DNA can be directly used in downstream applications without the need for further purification.

Storage and Stability

All components of the EZgene^TM Yeast gDNA Kit, except the Proteinase K,RNase A and Lyticase can be stored at 22^oC-25^oC and are guaranteed for at least 12 months from the dated of purchase. Once reconstituted in water,Proteinase K and Lyticase must be stored at -20^oC. Store RNase A at 4 ^oC. Under cool ambient conditions, a precipitate may form in the Buffer YBL/YTL. In case of such an event, heat the bottle at 37^oC to dissolve.  Store Buffer YTL/ YBL at room temperature.

Kit Contents

*Buffer YBL contains a chaotropic salt.

Before Starting

Please read the entire booklet to become familiar with the EZgene^TM Yeast gDNA Kit procedure.

•  Prepare a stock solution of Proteinase K (provided, 25 mg/mL) as follows and aliquot into adequate portions. Store aliquots at -20^oC.

•  Equilibrate Elution Buffer provided to 65^oC. Carry out all of centrifugation step at room temperature.
•  Dilute DNA Wash Buffer Concentrate with ethanol as follows:

Operating Protocol