Dear all professors, scholars and Drs.,
I am now facing a problem when I electroporate 15000 bp plasmid to lactobacillus casei atcc 393. I have tried several methods which involved:
Methods that I have tried (Not the steps)
1. 1% glycine treatment
2. Lithium acetate/DTT
3. 10 ug/ml ampicillin treatment
4. Spread plate on 2.5, 5, 10, 15 ug/ml erythromycin plate
5. 4 hrs recovery time
Even I use my positive control plasmid (7000bp) ----> can electroporate to others lacto strain, I cannot successfully electroporate the positive control into lacto atcc 393 strain.
Therefore, may I ask for your suggestions?
Is this related to the problem of the origin of replication (pAMBeta 1)?
Thank you.
Reading from your result about positive control, may I ask what was the electroporation procedure? One specie may have different electroporate condition and tolerance.
What type of cells are you using for electroporation? When doing this in the past with E. coli I have had to use a better cell line. Also, have you tried heat shock?
- Badges
- Science topic
- Similar topics
- Phytochemistry
- Extracts