Rehydration/Shampoo

Set up three baths for slides:

1. 400 ml Shampoo Soln, 50 °C
2. 400 ml H2O
3. 400 ml 95% Ethanol

• Mark slides on the back with diamond scribe pen.
• Expose slides to 65 mJ of UV irradiation.
• Dunk slides into Shampoo Solution at 50 °C, plunge for 15 seconds, incubate for 5 min.
• Transfer to 400 ml H2O for a few seconds to rinse
• Transfer to 400 ml 95% EtOH for a few seconds for drying
• Spin slides at 500 RPM, 5 min., Room Temperature!

Materials:	335 ml 1-methyl-2-pyrrolidinone
	5.5 g Succinic Anhydride
	15 ml 1M Na Borate pH 8 (premeasure into 15 ml conical)
	Rotating platform shaker
	Empty slide dish
	400 ml 95% EtOH bath
	Room Temperature H2O bath in beaker large enough to rotate slide rack

• Place 335 ml 1-methyl-2-pyrrolidinone into 500 ml beaker with stir bar
• Add 5.5 g Succinic anhydride
• When the last crystal dissolves add 15 ml 1M Na Borate pH 8.0 and transfer to a slide dish
• Plunge slide rack rapidly into blocking solution, plunge mix up and down for 30 seconds
• Shake gently on rotator for 15 minutes
• Drain blocking solution off slides and transfer to 50 °C water bath
• Incubate 60 seconds
• Transfer to 95% Ethanol bath to rinse
• Spin slides dry in the centrifuge, 500 RPM, 5 minutes, Room Temperature
• Store the slides in a dust free plastic slide box, they are ready for immediate use.