Koeris/Notebook/2007-1-15

Pseudomonas aeruginosa mer operon cloning 3

Preparation

What can I do in order to make sure the cloning works this time?

• Check concentrations of the chromatin DNA
• Do a dilution series over three orders of magnitude of the template DNA

PCR set-up of the operon

1. Master mix
   1. Template DNA equiv.
   2. ddH₂O to 50uL
2. Thermal profile
   1. 92 deg C - 5min
   2. 92 deg C - 30s
   3. 50 deg C - 45s
   4. 72 deg C - 120s
   5. 4 deg C - indefinite

Pipetting scheme

Column heading 1	Column heading 2	Column heading 3
Row heading 1	Cell 2	Cell 3
Row heading A	Cell B	Cell C

Gel image

1% agarose TAE gel, ran @ 100V for 60min