Resuspension solution for preparing brewer's yeast electrosensitive state cell and preparation method of brewer's yeast electrosensitive state cell

Abstract

The invention relates to a resuspension solution for preparing brewer's yeast electrosensitive state cells and a preparation method of the brewer's yeast electrosensitive state cells. The resuspensionsolution comprises 5-15mM of a glycine-NaOH buffer solution, ethylene glycol which accounts for 1-5% of the total volume of the resuspension solution, dimethyl sulfoxide which accounts for 2-8% of the total volume of the resuspension solution, and 0.5-1.5M of sorbitol. By adopting the resuspension solution, the problem that in the prior art brewer's yeast electrosensitive state cells cannot be cryopreserved is solved, the valence attenuation of the cryopreserved brewer's yeast electrosensitive state cells is alleviated, experiment requirements can be met, and the brewer's yeast electrosensitive state cells can be cryopreserved for a long time and do not need to be prepared and used on site.

Description

technical field [0001] The invention relates to a resuspension liquid used for preparing Saccharomyces cerevisiae electrotransfer competent cells and a preparation method of Saccharomyces cerevisiae electrotransfer competent cells. Background technique [0002] After treatment, foreign DNA can be taken up, and cells in this state are called "competent cells". In microbiology, molecular biology, genetic engineering and other research fields, it is often necessary to use various competent cells for DNA transformation operations. [0003] The preparation of competent cells is an important link in molecular biology research, and its quality directly affects the subsequent experimental work. Commonly used methods for preparing competent cells include calcium chloride method, Hanahan and Inoue method. The calcium chloride method is a conventional method for preparing competent cells in the laboratory. This method is simple and easy to implement, but the transformation efficiency...

AI Technical Summary

Problems solved by technology

The calcium chloride method is a conventional method for preparing competent cells in the laboratory. This method is simple and easy to implement, but the transformation efficiency of the prepared competent cells is low, which can only meet the needs of routine molecular biology experiments; , the degree of care and the purity of the reagents are high, and the repeatability is low; although the Inoue method has a high conversion efficiency of competent cells, it does not affect the OD of bacteria. 600 The value requirements are relatively strict, and the cultivation temperature is 18°C. Bacteria grow slowly at this temperature, the cultivation time is long, and the process is relatively cumbersome.

[0004] Moreover, the current yeast electrotransfer competent cells are ready-to-use, each preparation takes a lot of time and experience, and the completed electrotransfer competent cells lose their activity after freezing.