Team:Fatih Turkey/Notebook

14.07.11

PROBLEMS AND SOLUTIONS

# Prepare to competent materials;
*We prepared the competent materials for tomorrow
(tomorrow we will prepare the competents of b. subtilis and it will be 5th time:))
*we prepared our b subtilis plates for using tomorrow
*we have two plates for competent, one of them is wild type and other stain is ATCC6633
*we named them not to confuse, the ATCC663 strain's new name is Ahmet, wild type bacteria's new name is Mahmut :)
those names are turkish male names

# Transformation;
*we couldn't get the expected results at transformation of b. subtilis in this week today we will try to make the most succesful transformation ever :)
The Problems and The Solutions:
-We decided to some changeings at our procedure of competent
*incubation time will be a hour, firstly we will add the DNA to b. subtilis pellet without medium B then we will shake the mix for 30 min. and we  will add medium B on this mix than we will shake new mix again for 30 min.
*amount of DNA will be increased

SITUATION
Transformation; we prepared 4 plates for transformation
1. plate : b. subtilis pellet + 6 microliter plasmid ---> incubation/shaker30 min.--> adding 100 microliter medium B --->incubation/shaker30min. --->spreading 5ug/ml Chm plate
2. plate: b subtilis pellet + 10 microliter plasmid ---> incubation/shaker30 min.--> adding 100 microliter medium B --->incubation/shaker30min.----> spreading 5ug/ml  Chm plate
3. plate: b subtilis pellet + adding 100 microliter medium B ---> incubation/sahker 30 min.--->adding 10 microliter plasmid ---> incubation/shaker30min.----> spreading 5ug/ml Chm plate
4.plate: b subtilis pellet + adding 100 microliter medium B ---> incubation/sahker 30 min.--->adding 6 microliter plasmid ---> incubation/shaker30min.----> spreading 5ug/ml Chm plate