Y1

Buffer Y1 (yeast lysis buffer) used for the QIAGEN DNA extraction for yeast and QIAGEN Genomic-Tip DNA extraction protocol.

The buffer is used to digest the cell walls with a lytic enzyme (lyticase or Lallzyme MMX) after which protoplast are spinned down.

Work in a fume/safety hood (this buffer contains the highly toxic beta-mercaptoethanol)

Buffer Y1

• 1 M sorbitol
• 100 mM EDTA pH 8.0
• 14 mM β-mercaptoethanol

Buffer Y1 should always be stored at 4°C.

To make Y1:

• Dissolve 182.2 g sorbitol in 600 ml distilled water.
• Add 200 ml of a 0.5 M Na₂EDTA (pH 8.0) solution.
• Add 1 ml of β-mercaptoethanol (14.3 M).
• Adjust the volume to 1 liter with distilled water.
• pH does not need to be adjusted.