User:Tk/Notebook/MF-xfm/2008/04/27

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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
    • need 30
  • Primer
    • need 0.3 ul x 100 x 30 = 300 ul, we should have sufficient, but order another batch