This morning we continued with the gfp extraction. We put a resin/nickel mixture in a syringe. Then we put the gfp sample on top of the nuckel solution. The gfp should bind to the nickel. Tis is a time consuming process.

We aso took photos of our ps1 crystals. We had three sizes, micro, small and large. The large are big enough to be used in x-ray diffraction.
We also found that that our phlocyanin crystals are in Berkley and may be used on the syncchotron.

Jenniferand I worked with with Di on the GFP extraction. Di had previously centrifued the broth to extract the e-coli bacteria which had been kept an -80 C overnight. The cell walls of the baacteria was then broken using a sonicizer, sort of like a high tech sonic toothbrush. The mixture was then centrifued again. Tomorrow we will purify using a nickel column.

Also we carried on with our inorganic cystallisation. Our crystals of copper sulphate were excellent. We aslo created crystals of potassium chloride.

Jodie, Jannette and Petra were here unitl 9pm last night working on the power point. It looks excellent.

At this morning's seminar Craig gave an excellent power point presentation on the physics of photosynthesis and fluoresence. Craig was mostly concerned with the emitted light after PS1 have absorbed light. He studied this process with a smear camera which allows the amplitude and wavelength of a light pulse to be studied over a short period of time. Craig had found that here were four different light emissions with different time constants.

Next we talked with Petra about some possible inorganic crystal growing projects we could do next week which might be suitable for our classrooms. Also , next week, we will work on isolating the GFP protein.

This morning we took pictures of our lysozyme crystals. The we spent time in the cold room working n our PS1 crystals. WE had already planted small crystals inn a protein mix. There dialysis vessels had now created medium size crystals which were in the form of rods. These rod crystals were planted into another dialysis vessel with the correct buffer/salt concentration with the hope of creating large crystals

Today we continued our PCR lab. Our cultures had spent an hour in the PCR machine and had hopefully multiplied. Now we had to use a gel and electric voltage to see if that was true. We had created the gel yesterday. A sample of each PCR run with a loading dye was put into a small well at the top of the gell. We had 5 samples plus a standard. A volatge of 100V was placed across the gel and left for an hour. To see the DNA we had to add ethidium bromide which binds to the DNA. We then looked at the gel under a UV lamp. DNA had been multplied, the PCR had worked.

We also spent an hour with Professor Wachter discussing green fluorescent proteins(GFP). GFP are an immense area of interest. Nam is creating mutants of GFP to see if the can react in a faster amount, at present 1/2 hour

This morning we spent our time understanding PCR which is a method of copying parts of DNA in large quantities. Prof Rebekka Wachter gave us a introduction to PCR. It is very new technique. In short DNA is heated to break it into single strands(denaturation or melting). Then primers ( short strands of DNA with specific code) latch onto the single DNA strand(annealing). Then a special enzyme, DNApolymerase, which works at high temperature, links onto the DNA/primer complex and starts adding new amino acids. This cycle is repeated many cycles(30) and 2^30 or 10^9 new strands of DNA are created.
Nan, a graduate student, let us start a PCR lab.
Finally, with Petra Fromme, we put small seed crystals into our PS1 dialysis vessels.

This morning we spent the morning with Prof Petra Fromme preparing reaction vessels to create crystals of PS1. PS1 crystallisation uses dialysis to reduce the salt concentration to create crystals.
There were lots of different stages. We had to find the exact molarity of the protein. We did this by finding the molarity of the chlorophyll which we did by using a spectroanalyser.
We had to make dialysis tubes. The salt solution will pass through a membrane to a high salt conc leaving behind protein in a higher salt conc.