More Difficulties

I asked Brennan if he would like me to walk him to school on his second day of school and he said he would, so I saw him to the gate and then took the 8:40 bus to work. I started the column running again and stained the gel which supposedly contained the histones H3 and H4. It showed about the same thing as the previous day’s attempt, with the exception of a faint band in the H4 sample, running at about the same size as an unrelated protein that was contaminating the H3 sample. Kiyoe thought that this showed that the previous two attempts at purification had not been successful after all, and worried that something went wrong in the very first step when I prepared the DNA from the clone. If something had gone wrong at this step, all subsequent manipulations would have failed. She suggested that I start with the original sample we received from Tokyo and insert this into cells again. Meanwhile, I poured some gels in anticipation of running them once the column had finished. I got a fried chicken dish over rice with mayonnaise squirted over the top of it and rotini noodles in a tomato sauce for lunch. After the column finished running, I detected the amounts of protein using the colorimetric assay and ran a gel of the appropriate fractions. The results were a cause for concern: instead of two proteins in my purified complex, there were three! One of the proteins, H2A, looked like it had gone bad after being stored in the coldroom for about a month. The scale-up had failed; at least I had the very first complex formation attempt safely stored away.

I called before leaving work at 5:55 and asked the family to meet me at the bus stop so we could walk to the train station together to go to church. The service had been switched from Wednesday to Friday since Yudai, a brother who was visiting from Tokyo, would be preaching; this also meant that the Sunday service would be moved to Saturday in order to take advantage of his being in town. The family had luckily brought umbrellas, since it started pouring rain as soon as we were all assembled. Brennan had another rough day at school and had not enjoyed it any more that the first day. We took the train to Mikuni and ate dinner at the McDonalds that was located in the station. We then walked through the rain to church. Everyone there was excited to meet the family, and Yudai’s message was a good one- he continued the theme of the Holy Spirit that we had started the previous midweek service. They sang more than the usual number of songs in English, probably to help the family feel welcome. A number of us walked to the train station afterwards, Trudy walked ahead of the pack and talked with Nami, a sister she had hit it off with right away. Soon, the family was making its way across campus and had returned home by 10:30; luckily the rain had finally ended for the day.

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