cracking gel - MetabolicEngineeringGroupCBMA/MetabolicEngineeringGroupCBMA.github.io GitHub Wiki

http://stanxterm.aecom.yu.edu/wiki/index.php?page=Cracking_gel_method

Sucrose 25 g ddH2O 40 ml (Dissolve sucrose at 65 deg C) 5 M NaOH 5 mi 10% SDS 2.5 ml Add ddH20 to 50 ml final volume

alt.

Simply Cloning - Supplement 2 - Cracking Gel (https://www.youtube.com/watch?v=BfQ-UIabquk)

39.5 ml ddH20 2.5 mL 1M NaoH 2.5 mL SDS 10% 0.5mL 0.5M EDTA

Bacterial colonies of about 1mm size can directly be cracked right after picking trom the plate (no overnight culture needed). Pick the colony with a tooth-pick, and resuspend it in 20 µl LB medium or just water (by vigorously rotating and scraping the pick against wall of the tube).

Add cracking buffer to that 20 ul of bacterial suspension. Put the pick into a plastic tube with LB and antibiotics for overnight culture (or just inoculate an LB + antibiotic agar plate at specific position).