Transposon results
- Picked 177 colonies from 4/25 plates
- Picked into 96 well plates with 1.8 ml tet medium
- Regrew plates for more slow colonies
- Grew a full 96 well plate o/n
- Kept partially full plate for later growth
Scale up requirements
- Need about 2000-3000 transposon events
- about 20-30 plates worth
- We get 2 plates per 400 ul transform (as done 4/25)
- Need to do about 10 transformations, plan on 15
- We have sufficient transposome for 4 more transformations
- We have sufficient DNA for 5 additional transformations
- Need additional ME0/PCR DNA prep, 120 ng/ul column purified
- Need 12 x 12 plates (1161 + tet)
- Have 80, need 70; this is 4 sleeves or two liters of broth
- Plates
- Growth plate -- 2 ml deep well, sterile, need 30
- Sterile seals and lids for growth plates, need 30
- Each growth plate needs 200 ml of 1161 + tet broth or 6 liters
- Storage plates, 200 ul volume, need 30
- sterile seals and lids for storage plates, need 30
- Genomic preps
- Primer
- need 0.3 ul x 100 x 30 = 300 ul, we should have sufficient, but order another batch
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