[Note, we can make a 1L solution of the buffer minus lysozyme. Then, when we’re ready to extract a batch, we aliquot smaller volumes and then add the lysozyme. E.g. for 50 ml of buffer solution, we can add 200 mg of lysozyme.]
To make 1L of lysis buffer without lysozyme:
20 ml of 1M Tris-Cl (final concentration of 20mM Tris-Cl in buffer)
2 ml of 0.5M EDTA (final concentration of 2mM EDTA in buffer)
12ml Triton‱ X-100 (final concentration 1.2% Triton‱ X-100 in buffer).
200 mg of lysozyme powder/granules
Fill up to 50ml of the lysis buffer described above
*Adjust amount of each solution depending on amount of samples
Solution I (50mls) Enzymatic Lysis Buffer
20 mM Tris-HCl (pH 8) buffer
1.2% Triton X-100 detergent
1% SDS (diluted from 20% stock) 2.5mls (2500µl)
Glacial acetic acid 5.75mls