Amber #Entry 2 one of many failures and successes

Hellos,

The second week at my lab was exciting and reflecting. I became more independent at my work right now in terms of all the techniques and lab settings. Moreover, my mentor is starting to take classes starting from July, I will take more responsibility in proceeding her project. The gradual independence at my lab work also came with the price with failures and mistakes.

 My first task alone is to culture the LLCMK2 cells. The procedure can be simply broken down into washing the flask with PBS and adding medium and trypsin. The importance of splitting cells is to prevent the cells from clumping together. This is one of the basic techniques in a molecular biology lab since all the lab work will be based on using the cells. After watching the postdocs split the cells for a few times, I was prepared for this task. However, the bad news came on the weeknd when my mentor told me the cells actually died. Even though my mentor told it’s not a very important sample, I was frustrated because the tissue culture SEEMED so easy. Later we went over the steps again and concluded that I might pippet too hard on the walls that the cells died. It’s really hard to tell at that time because it’s impossible to visualize the cells without microscope. After this failure, I became extra careful with every single step at lab because any error can be amplified in the result. My next failure was ABRT. ABRT is using RNA as a template to make the cDNA. The procedure mainly consist of making the master mix of enzyme and buffer and dilute RNA. However, I put the primer without the ice in RT for 1 minute and that’s probably the reason it failed. I didn’t give that many thoughts at that time because I put them back onto ice again. Now I learned all these things I need to be extra careful with during my lab work. I feel really grateful of working at this lab because everyone here is so encouraging and friendly-- they are always willing to explain things for me and helping me find the mistakes.

  My accomplishment came at the success of my RNA extraction. After a week of practicing and learning, I did my own RNA extraction on Friday. The fact that my mentor said these samples will directly affect the grant due next Wednesday made me even more nervous. The 260/280 (a way to test the purity of extracted RNA) was really good and my mentor said I really helped her with the grant project. I was happy to hear the contribution I made to the lab.

Besides that, I am learning the program PRISM to process data derived from the qCR results. It mainly uses matrix and excel sheet to process the raw data. And I helped other postdocs with virus harvesting and gel extraction. Besides the work at lab, I started taking tennis lessons at UPenn tennis center. The food festival near spring garden was a lot of fun too.

 I am excited to continue my life at Philly and lab!