My apologies for the first post - It got away without the attachment.
Also sorry about the companies auto generated disclaimer.
I haven't seen commercial yeast mini-prep kits - but this modification
of Promega's Wizard miniprep kit has worked consistantly well for me.
My protocol was adapted from an "Promega Notes" article (sorry I lost
the reference) that they published a year or so ago. The lysis buffer
is stable for weeks at room temp.
Yeast MiniPrep with Wizard Mini-Prep purification
1. Grow 10 mL cultures overnight to stationary phase (selective media
2. Harvest cells by centrifugation, decant super, resuspend cells in
residual liquid and transfer to screw cap microfuge tube.
3. Pellet cells in microfuge, 20 seconds. Remove super, resuspend
cells in 200µl lysis buffer:
2% Triton X100
10mM Tris-Cl, pH 8
4. Add 200µl CHCl3/phenol/isoamyl alcohol, 24:24:1, 0.3 gm acid-washed
glass beads (0.5mm).
5. Vortex (high speed) 2 minutes.
6. Microfuge, 5 minutes.
7. Transfer super to clean microfuge tube, add 1 mL Promega Wizard
8. Process the resin as per the Wizard kit instructions (Transfer resin
to mini-column, apply vacuum to sample, wash column with wash
solution, spin put residual liquid from column, elute plasmid with warm
9. Use 5µl of mini-prep to transform 50µl of subcloning efficiency
E.coli (Gibco's DH5a or HB101) using the procedure provided with the
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