915
edits
Changes
Wrote initial protocol
==Materials and Buffers==
* Screw Capped Vials
* 30% KOH, prepared fresh
* 1M Sodium Sulfate
* Ethanol
* 50 mM Sodium Acetate, pH 4.8
* Amyloglucosidease
==Protocol==
# Weight out 30-90 mg tissue into screw cap vial and record weights.
# Add 300 uL 30% KOH and place on 95C heat block for 30 min with occasional mixing.
# Cool and then add 100 uL Sodium Sulfate and 800 uL Ethanol.
# Boil for 5 min.
# Centrifuge at 13 000 RPM for 5 min.
# Resuspend pellet in 200 uL water then add 400 uL ethanol. Boil 5 min, spin 5 min and Repeat wash steps twice more.
# Dry pellet
# Resuspend pellet in 200 uL of 50 mM Sodium Acetate, pH 4.8
# Add 0.3 mg/mL amyloglucosidase and place at 37C for 3h
# Quantify glucose
Reference:
PMID 15282316
* Screw Capped Vials
* 30% KOH, prepared fresh
* 1M Sodium Sulfate
* Ethanol
* 50 mM Sodium Acetate, pH 4.8
* Amyloglucosidease
==Protocol==
# Weight out 30-90 mg tissue into screw cap vial and record weights.
# Add 300 uL 30% KOH and place on 95C heat block for 30 min with occasional mixing.
# Cool and then add 100 uL Sodium Sulfate and 800 uL Ethanol.
# Boil for 5 min.
# Centrifuge at 13 000 RPM for 5 min.
# Resuspend pellet in 200 uL water then add 400 uL ethanol. Boil 5 min, spin 5 min and Repeat wash steps twice more.
# Dry pellet
# Resuspend pellet in 200 uL of 50 mM Sodium Acetate, pH 4.8
# Add 0.3 mg/mL amyloglucosidase and place at 37C for 3h
# Quantify glucose
Reference:
PMID 15282316
