1,973
edits
TOP10 chemically competent cells: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
TOP10 chemically competent cells (view source)
Revision as of 22:58, 8 November 2006
, 8 November 2006→Measurement of competence
| Line 59: | Line 59: | ||
* Incubate at 37 C for 1 hour in 2 ml centrifuge tubes rotated | * Incubate at 37 C for 1 hour in 2 ml centrifuge tubes rotated | ||
** using 2ml centrifuge tubes for transformation and regrowth works well because the small volumes flow well when rotated, increasing aeration. | ** using 2ml centrifuge tubes for transformation and regrowth works well because the small volumes flow well when rotated, increasing aeration. | ||
** For our plasmids (pSB1AC3, pSB1AT3) which are chloramphenicol and tetracycline resistant, we find growing for 2 hours yields many more colonies | |||
** Ampicillin and kanamycin appear to do fine with 1 hour growth | |||
* Plate 20 μl on AMP plates using 3.5 mm glass beads | * Plate 20 μl on AMP plates using 3.5 mm glass beads | ||
* Transformation efficiency is 15 x colony count x 10<sup>5</sup> | * Transformation efficiency is 15 x colony count x 10<sup>5</sup> | ||
