Team:Cambridge/LabBook/Week11

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Week 11: Monday 20th September - Sunday 26th September

Monday

107. Expt: Biobrick assembly of pBAD+Luc (PP + LC) and (Emily)

  • Plasmids already extracted
  • Restriction digest following protocol on p88 using these quantities (in µl):
pBAD PP Luc (1) PP Luc (2) LC Luc (1)
Nuclease-free H20 14 14 14 14
10x FD Buffer 2 2 2 2
Plasmid DNA 2 2 2 2
FD EcoRI 1 1 1 1
FD SpeI 1 0 0 0
FD XbaI 0 1 1 1
  • Gel electrophoresis
    • pBAD failed - band at ~5000bp, should be 1200
    • others look about right ~ between 3 & 5kb (should be 3600bp)
  • Gel extraction - results in -20°C freezer

108. Expt: Send off PP + pSB1C3, EPIC pBAD for sequencing (Emily and Peter)

  • Miniprep EPIC pBAD and nanodrop
  • Prepare correct concentrations (100ng/µl for plasmid, 3.2pmol/µl for primers) to be sent off


µl°C