User:Howard Boland/Notebook/Art from Synthetic Biology/2011/02/04

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Check Overnight Growth BBa_J04450

Growth was fine

Miniprep BBa_J04450

Eluted in 35µl Water

Double Digest BBa_J04450 E|P

Eluted in 35µl Water

Gel BBa_J04450 E|P

1% Agarose Gel, 04-FEBRURARY-2011-17:44 GMT

  1. Lane 1: 10µL 1kbp NEB Quick Ladder
  2. Lane 2: BLANK
  3. Lane 3: 50µL BBa_J04450 E|P (Expected 1069bp (part) and 5263bp backbone (pSB1A10))
  4. Lane 4-8: BLANK

04022011-backbone.jpg

Purified BBa_J04450 E|P

Purified Second Band from top, expected 5263bp backbone

Ligation BBa__katE E|S & BBa_I13504 X|P & BBa_J04450 E|P

Setup a ligation containing

  1. 3µl BBa_J04450 E|P
  2. 1µl BBa_I13504 X|P
  3. 1µl BBa__katE E|S
  4. 3µl Ligation Buffer
  5. 1µl Ligase
  6. 1.5µl ddH2O

Transformation of Ligation

The whole ligation was transformed in an carbenicillin dish