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

{| width="800"
 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] MF-xfm
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


 * colspan="2"|
 * colspan="2"|

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


 * }