Stocks from Bacillus Genetic Stock Center

Received a number of E. coli and Bacillus spp. from the Bacillus Genetic Stock Center in the mail on September 24, 2014. They have since been stored at 4C until then.

Today, media was created in order to inoculate broth cultures of the five E. coli strains.

E. coli Strains
ECE10 (MM294(pBS42))- on LB Cm5ug/ml
ECE32 (MC1061(pHP13)) - on LB Cm5ug/ml
ECE90 (TG1(pDG641)) - on LB Amp100ug/ml
ECE93 (TG1(pDG780)) - on LB Amp100ug/ml
ECE 98 (TG1(pDG1515))- on LB Amp100ug/ml

A single isolated colony (sometime multiple if they were small) from each E. coli were resuspended in 10mL of LB broth +Abx and incubated at 37C at 225 rpm.

Antibiotic broth media

Stock Ampicillin was at 100 mg/ml. So, 10 ul was added to 10 mL of LB broth in three 50 ml conical tubes.

Stock Chloramphenicol was at 50 mg/ml. So, 2.5 ul was added to 10 mL of LB broth in two 50 ml conical tubes.

E. coli B/r was subsequently inoculated in Abx media as well to serve as a negative control.

Cultures will incubate overnight and freezer stocks will be made.

There were a number of B. spp. also included with the order, however the media required to re-suspend the spores is not prepared yet and will have to wait until a later date.

//EWW