From OpenWetWare
< User:Tk‎ | Notebook‎ | MF‎ | 2008‎ | 11
Owwnotebook icon.png Mesoplasma florum simplification <html><img src="/images/9/94/Report.png" border="0" /></html> Main Page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Part construction

  • Miniprep of four cultures each for J70027, J70024, J70025
  • Cut J70027, J70025 with E-S
  • Cut J70024 with X-P
  • Ran 5 ul on an E-Gel
  • Lanes: marker, J70027ABC, J70024ABC, J70025ABC, J70027D, marker


Choose J70027A, J70024A, J70025A

Assemble Tet cassette

J70028 (Kan) = J70025 (CM) + J70024 (CM): this should be tet resistant also

  • Four versions, J70025A/C with J70024A/C
    • AA, AC, CA, CC

Sequence new components

Sequence with VF2, VR:

  • J70027A,D
  • J70024A,C
  • J70025A,C