Hi i am planning to do bacterial transformations for protein expression of two different types but due to the plates being over a month old and having no other sources to inoculate i am forced to use two month old plates.
I plan on creating plate transformations ultimately into E.coli BL- 21 for my protein expression. To go about this by getting mini-prep DNA and then setting up a transformation
However due to the age of the current colonies should i do it first into E.coli XL1 Blue which is good for replication numbers
or should i just go straight into E.coli BL-21
In my experience there is very low yield of plasmid DNA if make a preparation from BL21 strain. I suggest firstly transform any XL1, XL10 or top10 strains.
Hi there,
BL21 strain is genetically unstable therefore the best for storage of the plasmid is either plasmid prep or XL1 transformed strain (or any other cloning strains which are genetically much more stable).
- Agree with Airit
- Make your plasmid prep from XL-1 strain then re transform into BL-21 for protein expression purposes
Hi,
They are exactly right. first transform it in XL-1 then in BL21.
GOOD LUCK:)
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