User:NYOUSHA Yousefi/Notebook/Protein Engineering/2013/02/05
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Hb buffer exchange   The wild type Hb's buffer was exchanged to
a new buffer containing 1g manitol, 0.5g PEG, and 50ml phosphate
pH 7  Procedure 1    1. Column chromatogrophy was rinsed with
25ml of distilled water and 25ml of the new buffer solution
containing 0.5g PEG, 1g manitol,and 50ml of phosphate buffer, pH
8   2. 2.5ml of WT Hb and 2.5ml of the new buffer were added to
the column and this 5ml was collected in a new tube (2% manitol,
1% PEG, WT Hb, pH 7 phosphate)  Objective 2  Extracting bacteria
cells  Procedure 2    1. The bacteria expressing Mn and Ni were
allowed to grow in media for 6 hours after addition of IPTG,
which is a signal that stimulates protein expression   2. The
media was d