Kirsty the Scientist

As I was talking about earlier this week, I've often been running more than one experiment at the same time during a week. But today, the differences between the two experiments seemed quite striking!  The two experiments are to look for two different indicators to suggest that my protein of interest is involved in making my cell type increase during cell stress/inflammation:  1. Look for the protein itself. This involves separating the proteins using electrophoresis (pulling the proteins through a polyacrylamide gel using an electric current to draw the proteins through the tiny pores of the bell - the smaller proteins get to the bottom of the gel quicker as they can wiggle through the holes faster. Then using a method called Wet Transfer, which pulls the protein out of the gel and onto a PVDF membrane - a technique that uses litres of buffer.  2. Looking for the genes that are expressed - suggesting that the cell is producing the proteins and using the part of the...

Similar to yesterday, (and most days nowadays!) I was working on two goals:  1. Prep more cells for use for a new experiment for this week.  2. Detect the secondary antibody signals using the Western blots using two slightly different methods (which I can compare to see which is better). Li-Cor detects the signals using fluorescence using a shiny new machine the institute has; whereas ECL is the standard method which uses light.  We also had our first full group meeting today, where the research team gets together and we can discuss the results of everyone's experiments - which will now become a bit more of a regular occurrence! 

Still processing the information gathered from last week's experiments (results collected on Friday). Reprobing the Western blots with more antibody to compare the two proteins we're looking for.  Also learnt how to use an 'alternative Excel' today - called GraphPad Prism, which is a bit better than Excel for producing the lovely good-looking diagrams and graphs ready for publication and to pop in my dissertation.  All in all, these first results make a good start to the project, and has given a few more clues as to the direction this project is going to take over the next couple of weeks. 

This evening I came home and almost instantly fell asleep! It's been a busy week, and a lot of information to take in each day and new things to learn! And today's results were a little anti-climatic, with some contamination in one of the lanes of the western blot, and the PCR results still a bunch of numbers that I'll be compiling into a graph over the weekend before they make much sense! 

Another busy day, preparing the plates of cells to be tested in 2 different experiments - one to search for the protein itself and one looking for the DNA that makes the protein.  On the cusp of finding out the results of a week's worth of work, I was reminded of something I listened to in a podcast today. It was a quote from Richard Feynman, the renowned physicist: "The scientist has a lots of experience with ignorance, doubt and uncertainty., and this experience is of very great importance. When a scientist doesn't know the answer to a problem they are ignorant, when they have a hunch as to what a result is they are uncertain, and when they are pretty darn sure of the result they are in doubt. Science is a satisfactory philosophy where doubt is not to be feared but welcomed and discussed." "To paraphrase - s cience is the enemy of the certain and being shown to be wrong is an invaluable part of learning about nature." And that's something I will k...

This morning we headed across town with a box of dry ice to collect the cells from the liquid nitrogen store. We took them back to tissue culture to be defrosted, resurrected and ready to grow over the next few days. Also finished off the Western blot today, so should get some good results from those tomorrow!  Although it's been less than a week so far, I'm really enjoying working with and getting on really well with everyone in my research group. Now we're actually getting on with some proper lab work, I'm so excited for the next few weeks!