Anybody know?

Assalamualaikum........

Today is the hundreds time after i tried so many time to get the effect of different pH on my protein solubility..as was predicted by me and my supervisor...the solubility of my protien may increase when the pH during fermentation is far away from pI value of my protein. But....what was expected....just as an expectation...... "harapan tinggal harapan... =("....

From the first i tried to prepare the buffer for each of pH until the protein quantification........still dissappointed...How difficult to prepare the buffer (what i had faced before......waiting for the stock..looking for the chemical...searching for the methodology.....calculating using Handerson-hasselbach......)----> actually not as difficult as what had been happen to me. That remind me about "sign"....sign is situation in which when we want to do something, the starting job reflect to the final job or the results. that is what "org2 tua" said...when you feel something uncomfortable b4 doing something,dont proceed?...sometimes good to follow this =)....hmmm....out of topic.its ok, at least i can release all the blocking2 area in my mind..

Back to the topic, anybody can help me,give some idea to make my project succesfull...i want to see the effect of different pH on my protein solubility. At the same time looking for the pH that give higher solubility. im using commercial media...TB(terrific broth),in TB media, the buffer system is phosphate buffer (0.06M), i increased the buffering capacity into 0.2M at the same time adjust the pH from 6 to 8 at the starting point of induction. Before this, i tried to adjust the pH from the starting time of fermentation,but the result is very2 frustrated =(.... all the cells in prepared buffered-media dead!!!....sob!sob!..

Then...i tried using TB for all pH,then once induce,change the pH with buffer of different pH. Still...the growth of e.coli not as expected....very slow....compared to the control....Now, im very tired to do all these. The latest trial also, i dont have any intention to proceed with the next step of protein extraction.

Whar r your suggestion?? help me......