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