Tag Info

New answers tagged

1

Tryptone is hydrolysate of casein- a milk protein- and may contain residual lactose as impurity. It is one of the constituent LB media. Addition of glucose may prevent induction of the cloned gene under lac promoter (or T7 polymerase under lac promoter in case of T7 promoter vectors) by catabolic repression. Induction of protein production is not desirable. ...


10

This is a matter of pragmatism in the culture process. Taking 100 colonies instead of 1 increases the inoculation volume by a factor of 100, which then saves you perhaps 2 hours of bacterial growth time before your culture reaches the OD you want. However, mutations and loss of plasmid in culture, while unlikely, are possible, especially if the bacteria ...


4

While all colonies on a selective media SHOULD contain a resistance gene, that's a long way from them containing the plasmid you want. A typical cloning experiment will insert a gene of interest into a empty vector backbone to make an intact plasmid. The backbone usually contains some antibiotic resistance gene. If your gene of interest is not correctly ...



Top 50 recent answers are included