2/13/24

Dear Aryan,

You cannot use lysozyme to digest yeast cell walls. Lysozyme attacks the peptidoglycan of gram-positive bacterial cell walls. The fabric of peptidoglycan is very different from that of the yeast cell wall, which is made mainly of glucan and mannan, plus some chitin. Lysozyme will not touch this.

I used zymolyase a number of times and it works very well. It requires dithiothreitol (DTT) to break disulfide bonds in the yeast cell wall to facilitate lysis by the enzyme. Cell lysis can be accomplished in ~an hour or less If you don't have zymolyase in your lab, you may have to purchase it. It is available from Zymo Research in Tuftin, California, USA. This is their website: https://www.zymoresearch.com/pages/all-products.

I hope this information helps you. Good luck with your research.

Bill Colonna Dept. Food Science & Human Nutrition, Iowa State University, Ames, Iowa, USA [email protected]

Hi Aryan,

Hope you are doing well. Lysozyme will not help as mentioned by @ W.J. Colonna. Maybe you could try the method used in the article attached below. The protocol involves lysis of yeast cells from liquid culture in a lithium acetate (LiOAc)–SDS solution. This method has been used to extract genomic DNA from yeasts for PCR-based applications. The paper has recommended the use of 200 mM LiOAc and 1% SDS in the lysis solution and carrying out the lysis at 70°C for ≥ 5 min. You may modify the protocol based on your requirement (Comet assay).

You may want to refer to the article and supplementary material attached below.

Article Extraction of genomic DNA from yeasts for PCR-based applications

https://www.future-science.com/doi/suppl/10.2144/000113672/suppl_file/btn5005-bm-kristjuh_160476a.pdf

Hope this is helpful!

Best Wishes,

Malcolm Nobre

(Ex-Xavierite, Bombay)

Thank you so much W.J. Colonna and Malcolm Nobre for your responses.