So, after getting some food in my belly and some sleep I have been able to return to the lab much stronger and focused. I can proudly say that I am out frustration alley.
I have decided that I WILL NOT BE DEFEATED. Research is filled with up days and down days and yesterday was my down day. It has become clear to me that I have chosen this path and I will stick with it. The joy truly comes after failing so many times and finally succeeding.
With much praying, reading and hoping. I think I’ve been able to figure out what I did wrong. I used way too little primary antibody. Today I took a plunge head first into rerunning my experiment. I have separated my proteins and transferred them to my nitrocellulose membrane. I have checked to see that my proteins transferred with Ponceau stain and it did. Now, I’m blocking. I have decided to block my membrane for 40 minutes, incubate membrane in primary antibody for an hour, wash membrane 3 times for 5 minutes each time and incubate in my secondary antibody for 30 to 45 minutes. It was brought to my attention that 70% of the primary binds to my specified protein within the first hour. Let me see how well this works.
My protein of interest today is P53 as my title suggests and come rain or shine, I WILL find it. This protein has eluded me for long enough. It has laughed while I cried, it has hidden while I searched and today is the day out paths will finally cross and when this happens, I will very gladly put up a blog with the title “VICTORY IS MINE” but I wont rejoice just yet.
All this searching and questing is being done less than a day before my presentation. Tomorrow morning, I’ll be presenting an introduction of my thesis project. What I intend to do, what I have done so far, etc. Long story short, I’m nervous and excited and nervous and hopeful and..oh wait did I mention I was nervous? I know my research, I know my hypothesis, I know my background information. I have read quite a bit but I feel like I don’t know anything. I feel like i need to read some more journals. One of my wonderful professors has told me that being nervous should make my brain more alert and I hope this will be the case.
Anyway, look out for my “VICTORY IS MINE” post…..(fingers crossed)
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