SynSCOBY Part 2 Growing Media E.coli

 October 25

Growing critters ins 2 media (LB, YPD)

• PNC (e.coli, plasmid  : LB. 37C
• PNS (e.coli, plasmid) : LB 37C
• BY4741(yeast) : YPD             30c
• VENUS (uria) (yeast): YPD.  30c
• 
  

Pour from the stock to a separate tube is done in the sterile hood 

Streaking from Plates. with the microbes from (Part1) : 

Taking a loopful of microbe culture (a colony) from the plates and putting them in Media

All Microbes grown overnight in Liquid Media

• PNC (e.coli, plasmid  : LB. 37C
• PNS (e.coli, plasmid) : LB 37C
• BY4741(yeast) : YPD             30c
• VENUS (uria) (yeast): YPD.  30c

For all microbes make glycerol stock 

60% glycerol to start and mix 50/50 with microbe culture. 30% Final

Store at -80C

JUST FOR E.COLI - 

Using LB + KAN

KAN is only for E.coli - use at 50-ug/mL

*Storing at -20oC and avoiding repeated freeze/thaw cycles will keep most antibiotics viable for at least 6 months.

Adding an antibiotic resistance gene to the plasmid solves both problems at once – it allows a us to easily detect plasmid-containing bacteria when the cells are grown on selective media, and provides those bacteria with a pressure to keep the plasmid

October 26

After being left overnight E.Coli s

YEAST ONLY

For all microbes make glycerol stock 

60% glycerol to start and mix 50/50 with microbe culture. 30% Final

Store at -80C

GROWS IN YPD in 30C

is placed on the 30oC 

Next Day (October 26)